Inhibitors of Akt activity

ABSTRACT

Invented are novel pyrazole compounds, the use of such compounds as inhibitors of protein kinase B activity and in the treatment of cancer and arthritis.

This application is a 371 of International Application No.PCT/US2008/058603, filed 28 Mar. 2008, which claims the benefit of U.S.Provisional Application No. 60/908,741, filed 29 Mar. 2007.

FIELD OF THE INVENTION

This invention relates to novel pyrazole compounds, the use of suchcompounds as inhibitors of protein kinase B (hereinafter PKB/Akt, PKB orAkt) activity and in the treatment of cancer and arthritis.

BACKGROUND OF THE INVENTION

The present invention relates to 1H-imidazo[4,5-c]pyridin-2-ylcontaining compounds that are inhibitors of the activity of one or moreof the isoforms of the serine/threonine kinase, Akt (also known asprotein kinase B). The present invention also relates to pharmaceuticalcompositions comprising such compounds and methods of using the instantcompounds in the treatment of cancer and arthritis (Liu et al. CurrentOpin. Pharmacology 3:317-22 (2003)).

Apoptosis (programmed cell death) plays essential roles in embryonicdevelopment and pathogenesis of various diseases, such as degenerativeneuronal diseases, cardiovascular diseases and cancer. Recent work hasled to the identification of various pro- and anti-apoptotic geneproducts that are involved in the regulation or execution of programmedcell death. Expression of anti-apoptotic genes, such as Bcl2 orBcl-x_(L), inhibits apoptotic cell death induced by various stimuli. Onthe other hand, expression of pro-apoptotic genes, such as Bax or Bad,leads to programmed cell death (Adams et al. Science, 281:1322-1326(1998)). The execution of programmed cell death is mediated by caspase-1related proteinases, including caspase-3, caspase-7, caspase-8 andcaspase-9 etc (Thornberry et al. Science, 281:1312-1316 (1998)).

The phosphatidylinositol 3′-OH kinase (PI3K)/Akt/PKB pathway appearsimportant for regulating cell survival/cell death (Kulik et al. Mol.Cell. Biol. 17:1595-1606 (1997); Franke et al, Cell, 88:435-437 (1997);Kauffmann-Zeh et al. Nature 385:544-548 (1997) Hemmings Science,275:628-630 (1997); Dudek et al., Science, 275:661-665 (1997)). Survivalfactors, such as platelet derived growth factor (PDGF), nerve growthfactor (NGF) and insulin-like growth factor-1 (IGF-I), promote cellsurvival under various conditions by inducing the activity of PI3K(Kulik et al. 1997, Hemmings 1997). Activated PI3K leads to theproduction of phosphatidylinositol (3,4,5)-triphosphate (PtdIns(3,4,5)-P3), which in turn binds to, and promotes the activation of, theserine/threonine kinase Akt, which contains a pleckstrin homology(PH)-domain (Franke et al Cell, 81:727-736 (1995); Hemmings Science,277:534 (1997); Downward, Curr. Opin. Cell Biol. 10:262-267 (1998),Alessi et al., EMBO J. 15: 6541-6551 (1996)). Specific inhibitors ofPI3K or dominant negative Akt/PKB mutants abolish survival-promotingactivities of these growth factors or cytokines. It has been previouslydisclosed that inhibitors of PI3K (LY294002 or wortmannin) blocked theactivation of Akt/PKB by upstream kinases. In addition, introduction ofconstitutively active PI3K or Akt/PKB mutants promotes cell survivalunder conditions in which cells normally undergo apoptotic cell death(Kulik et al. 1997, Dudek et al. 1997).

Analysis of Akt levels in human tumors showed that Akt2 is overexpressedin a significant number of ovarian (J. Q. Cheung et al. Proc. Natl.Acad. Sci. U.S.A. 89:9267-9271(1992)) and pancreatic cancers (J. Q.Cheung et al. Proc. Natl. Acad. Sci. U.S.A. 93:3636-3641 (1996)).Similarly, Akt3 was found to be overexpressed in breast and prostatecancer cell lines (Nakatani et al. J. Biol. Chem. 274:21528-21532(1999). It was demonstrated that Akt-2 was over-expressed in 12% ofovarian carcinomas and that amplification of Akt was especially frequentin 50% of undifferentiated tumors, suggestion that Akt may also beassociated with tumor aggressiveness (Bellacosa, et al., Int. J. Cancer,64, pp. 280-285, 1995). Increased Akt1 kinase activity has been reportedin breast, ovarian and prostate cancers (Sun et al. Am. J. Pathol. 159:431-7 (2001)).

The tumor suppressor PTEN, a protein and lipid phosphatase thatspecifically removes the 3′ phosphate of PtdIns(3,4,5)-P3, is a negativeregulator of the PI3K/Akt pathway (Li et al. Science 275:1943-1947(1997), Stambolic et al. Cell 95:29-39 (1998), Sun et al. Proc. Natl.Acad. Sci. U.S.A. 96:6199-6204 (1999)). Germline mutations of PTEN areresponsible for human cancer syndromes such as Cowden disease (Liaw etal. Nature Genetics 16:64-67 (1997)). PTEN is deleted in a largepercentage of human tumors and tumor cell lines without functional PTENshow elevated levels of activated Akt (Li et al. supra, Guldberg et al.Cancer Research 57:3660-3663 (1997), Risinger et al. Cancer Research57:4736-4738 (1997)).

These observations demonstrate that the PI3K/Akt pathway plays importantroles for regulating cell survival or apoptosis in tumorigenesis.

Three members of the Akt/PKB subfamily of second-messenger regulatedserine/threonine protein kinases have been identified and termedAkt1/PKBα, Akt2/PKBβ, and Akt3/PKBγ respectively. The isoforms arehomologous, particularly in regions encoding the catalytic domains.Akt/PKBs are activated by phosphorylation events occurring in responseto PI3K signaling. PI3K phosphorylates membrane inositol phospholipids,generating the second messengers phosphatidyl-inositol3,4,5-trisphosphate and phosphatidylinositol 3,4-bisphosphate, whichhave been shown to bind to the PH domain of Akt/PKB. The current modelof Akt/PKB activation proposes recruitment of the enzyme to the membraneby 3′-phosphorylated phosphoinositides, where phosphorylation of theregulatory sites of Akt/PKB by the upstream kinases occurs (B. A.Hemmings, Science 275:628-630 (1997); B. A. Hemmings, Science 276:534(1997); J. Downward, Science 279:673-674 (1998)).

Phosphorylation of Akt1/PKBα occurs on two regulatory sites, Thr³⁰⁸ inthe catalytic domain activation loop and on Ser⁴⁷³ near the carboxyterminus (D. R. Alessi et al. EMBO J. 15:6541-6551 (1996) and R. Meieret al. J. Biol. Chem. 272:30491-30497 (1997)). Equivalent regulatoryphosphorylation sites occur in Akt2/PKBβ and Akt3/PKBγ. The upstreamkinase, which phosphorylates Akt/PKB at the activation loop site hasbeen cloned and termed 3′-phosphoinositide dependent protein kinase 1(PDK1). PDK1 phosphorylates not only Akt/PKB, but also p70 ribosomal S6kinase, p90RSK, serum and glucocorticoid-regulated kinase (SGK), andprotein kinase C. The upstream kinase phosphorylating the regulatorysite of Akt/PKB near the carboxy terminus has not been identified yet,but recent reports imply a role for the integrin-linked kinase (ILK-1),a serine/threonine protein kinase, or autophosphorylation.

Inhibition of Akt activation and activity can be achieved by inhibitingPI3K with inhibitors such as LY294002 and wortmannin. However, PI3Kinhibition has the potential to indiscriminately affect not just allthree Akt isozymes but also other PH domain-containing signalingmolecules that are dependent on PdtIns(3,4,5)-P3, such as the Tec familyof tyrosine kinases. Furthermore, it has been disclosed that Akt can beactivated by growth signals that are independent of PI3K.

Alternatively, Akt activity can be inhibited by blocking the activity ofthe upstream kinase PDK1. The compound UCN-01 is a reported inhibitor ofPDK1. Biochem. J. 375(2):255 (2003). Again, inhibition of PDK1 wouldresult in inhibition of multiple protein kinases whose activities dependon PDK1, such as atypical PKC isoforms, SGK, and S6 kinases (Williams etal. Curr. Biol. 10:439-448 (2000).

Small molecule inhibitors of Akt are useful in the treatment of tumors,especially those with activated Akt (e.g. PTEN null tumors and tumorswith ras mutations). PTEN is a critical negative regulator of Akt andits function is lost in many cancers, including breast and prostatecarcinomas, glioblastomas, and several cancer syndromes includingBannayan-Zonana syndrome (Maehama, T. et al. Annual Review ofBiochemistry, 70: 247 (2001)), Cowden disease (Parsons, R.; Simpson, L.Methods in Molecular Biology (Totowa, N.J., United States), 222 (TumorSuppressor Genes, Volume 1): 147 (2003)), and Lhermitte-Duclos disease(Backman, S. et al. Current Opinion in Neurobiology, 12(5): 516 (2002)).Inhibition of Akt has also been implicated in the treatment ofleukemias, (J. C. Byrd, S. Stilgenbauer and I. W. Flinn “Chroniclymphocytic leukemia.” Hematology/the Education Program of the AmericanSociety of Hematology. American Society of Hematology. Education Program(2004), 163-83). Akt3 is up-regulated in estrogen receptor-deficientbreast cancers and androgen-independent prostate cancer cell lines andAkt2 is over-expressed in pancreatic and ovarian carcinomas. Akt1 isamplified in gastric cancers (Staal, Proc. Natl. Acad. Sci. USA 84:5034-7 (1987) and upregulated in breast cancers (Stal et al. BreastCancer Res. 5: R37-R44 (2003)). Therefore a small molecule Akt inhibitoris expected to be useful for the treatment of these types of cancer aswell as other types of cancer. Akt inhibitors are also useful incombination with further chemotherapeutic agents.

It is an object of the instant invention to provide novel compounds thatare inhibitors of Akt/PKB.

It is also an object of the present invention to provide pharmaceuticalcompositions that comprise a pharmaceutical carrier and compounds usefulin the methods of the invention.

It is also an object of the present invention to provide a method fortreating cancer that comprises administering such inhibitors of Akt/PKBactivity.

It is also an object of the present invention to provide a method fortreating arthritis that comprises administering such inhibitors ofAkt/PKB activity.

SUMMARY OF THE INVENTION

This invention relates to novel compounds of Formula (I):

wherein:

-   -   Q is selected from: phenyl, substituted phenyl, benzyl, and        benzyl wherein the aromatic ring is substituted;    -   R¹ is selected from: hydrogen, trifluoromethyl, hydroxy,        —C₁-C₂alkyl, and halogen;    -   L is selected from: nitrogen and —C(H)—;    -   P is selected from: nitrogen and —C(R⁴⁰)—, where R⁴⁰ is selected        from: hydrogen, —C₁-C₄alkyl, and halogen;    -   A is selected from: —C(O)— and —N(H)—;    -   B is selected from: —C(O)— and —N(H)—; and    -   X, Y and Z are independently selected from: nitrogen, —C(H)—,        and —C(R²)—, wherein R² is selected from halogen,        trifluoromethyl, hydroxy, and —C₁-C₄alkyl;        or a pharmaceutically acceptable salt thereof;    -   provided:    -   A and B are not the same; and    -   provided:    -   X, Y and Z are not all nitrogen; and    -   provided:    -   that at most one of P and L are nitrogen.

This invention relates to novel compounds of Formula (II):

wherein:

-   -   Q is selected from: phenyl, substituted phenyl, benzyl, and        benzyl wherein the aromatic ring is substituted;    -   R¹ is selected from: hydrogen, trifluoromethyl, hydroxy,        —C₁-C₂alkyl, and halogen;    -   R³ is selected from: hydrogen, trifluoromethyl, hydroxy,        —C₁-C₂alkyl, and halogen;    -   L is selected from: nitrogen and —C(H)—;    -   P is selected from: nitrogen and —C(R⁴⁵)—, where R⁴⁵ is selected        from: hydrogen, —C₁-C₄alkyl, and halogen;    -   A is selected from: —C(O)— and —N(H)—;    -   B is selected from: —C(O)— and —N(H)—; and    -   X and Y are independently selected from: nitrogen, —C(H)—, and        —C(R²)—, wherein R² is selected from halogen, trifluoromethyl,        hydroxy, and —C₁-C₄alkyl;        or pharmaceutically acceptable salt thereof;    -   provided:    -   A and B are not the same; and    -   provided:    -   that at most one of P and L are nitrogen.

This invention relates to a method of treating cancer, which comprisesadministering to a subject in need thereof an effective amount of anAkt/PKB inhibiting compound of Formula (I) or Formula (II).

This invention relates to a method of treating arthritis, whichcomprises administering to a subject in need thereof an effective amountof an Akt/PKB inhibiting compound of Formula (I) or Formula (II).

The present invention also relates to the discovery that the compoundsof Formula (I) and Formula (II) are active as inhibitors of Akt/PKB.

In a further aspect of the invention there is provided novel processesuseful in preparing the presently invented Akt/PKB inhibiting compounds.

Included in the present invention are pharmaceutical compositions thatcomprise a pharmaceutical carrier and compounds useful in the methods ofthe invention.

Also included in the present invention are methods of co-administeringthe presently invented Akt/PKB inhibiting compounds with further activeingredients.

DETAILED DESCRIPTION OF THE INVENTION

This invention relates to compounds of Formula (I) and Formula (II) asdescribed above.

The presently invented compounds of Formula (I) and Formula (II) inhibitAkt/PKB activity. In particular, the compounds disclosed herein inhibiteach of the three Akt/PKB isoforms.

Included among the presently invented compounds of Formula (I) arecompounds of Formula (IA):

wherein:

-   -   Q is selected from: phenyl, substituted phenyl, benzyl, and        benzyl wherein the aromatic ring is substituted;    -   R¹ is selected from: hydrogen, —C₁-C₂alkyl, and halogen;    -   R⁴ is selected from: hydrogen, —C₁-C₄alkyl, and halogen;    -   A is selected from: —C(O)— and —N(H)—;    -   B is selected from: —C(O)— and —N(H)—; and    -   X, Y and Z are independently selected from: nitrogen, —C(H)—,        and —C(R²)—, wherein R² is selected from halogen and        —C₁-C₄alkyl;        or a pharmaceutically acceptable salt thereof;    -   provided:    -   A and B are not the same; and    -   provided:    -   X, Y and Z are not all nitrogen.

Included among the presently invented compounds of Formula (II) arecompounds of Formula (IIA):

wherein:

-   -   Q is selected from: phenyl, substituted phenyl, benzyl, and        benzyl wherein the aromatic ring is substituted;    -   R¹ is selected from: hydrogen, —C₁-C₂alkyl, and halogen;    -   R³ is selected from: hydrogen, —C₁-C₂alkyl, and halogen;    -   A is selected from: —C(O)— and —N(H)—;    -   B is selected from: —C(O)— and —N(H)—; and    -   X and Y are independently selected from: nitrogen, —C(H)—, and        —C(R²)—, wherein R² is selected from halogen and —C₁-C₄alkyl;        or a pharmaceutically acceptable salt thereof;    -   provided:    -   A and B are not the same.

Included among the presently invented compounds of Formula (I) andFormula (II) are:

-   N-[2-amino-1-(phenylmethyl)ethyl]-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-6-(1-methyl-1H-pyrazol-5-yl)-3-pyridinecarboxamide;-   N-(2-amino-1-benzylethyl)-5-(1-methyl-1H-pyrazol-5-yl)pyrimidine-2-carboxamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-2-chloro-4-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-3-chloro-4-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-pyrazol-5-yl)-3-(trifluoromethyl)benzamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-chloro-4-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-4-chloro-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-6-chloro-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-6-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-5-(1-methyl-1H-pyrazol-5-yl)-3-pyridinecarboxamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-2-(1-methyl-1H-pyrazol-5-yl)-4-pyridinecarboxamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-3-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-3-methyl-4-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-methyl-4-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-3-hydroxy-4-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-3-chloro-5-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-2-chloro-5-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-4-chloro-3-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-chloro-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-chloro-3-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-3-fluorobenzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-3-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)benzamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluoro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)benzamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)benzamide;-   N-[2-amino-1-(phenylmethyl)ethyl]-3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzamide;-   3-amino-N-[3-chloro-4-(1-methyl-1H-pyrazol-5-yl)phenyl]-2-phenylpropanamide;-   3-amino-N-[3-chloro-4-(1-methyl-1H-pyrazol-5-yl)phenyl]-2-(phenylmethyl)propanamide;-   3-amino-N-[3-(1-methyl-1H-pyrazol-5-yl)phenyl]-2-(phenylmethyl)propanamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide;-   N-[(1S)-2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide;-   N-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide;-   N-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide;-   N-[(1S)-2-amino-1-(phenylmethyl)ethyl]-3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide;-   N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(4-chloro-1-methyl-1H-1,2,3-triazol-5-yl)benzamide;-   N-[(1S)-2-amino-1-(phenylmethyl)ethyl]-4-(4-chloro-1-methyl-1H-1,2,3-triazol-5-yl)benzamide;-   N-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-4-(4-chloro-1-methyl-1H-1,2,3-triazol-5-yl)benzamide;-   N-[(1S)-2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide;    and-   N-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide;    or a pharmaceutically acceptable salt thereof.

Compounds of Formula (I) and Formula (II) are included in thepharmaceutical compositions of the invention and used in the methods ofthe invention.

Certain of the compounds described herein may contain one or more chiralatoms, or may otherwise be capable of existing as two enantiomers.Accordingly, the compounds of this invention include mixtures ofenantiomers as well as purified enantiomers or enantiomerically enrichedmixtures. Also, it is understood that all tautomers and mixtures oftautomers are included within the scope of the compounds of Formula (I)and Formula (II).

Certain compounds described herein may form a solvate which isunderstood to be a complex of variable stoichiometry formed by a solute(in this invention, a compound of Formula (I) or a salt thereof, or acompound of Formula (II) or a salt thereof) and a solvent. Such solventsfor the purpose of the invention may not interfere with the biologicalactivity of the solute. Examples of suitable solvents include, but arenot limited to, water, methanol, ethanol and acetic acid. Preferably thesolvent used is a pharmaceutically acceptable solvent. Examples ofsuitable pharmaceutically acceptable solvents include, withoutlimitation, water, ethanol and acetic acid.

By the term “aryl”, and derivatives thereof, used alone or as part of alarger moiety as in “—(CH₂)_(m)aryl” as used herein, unless otherwisedefined, is meant monocyclic, bicyclic, and tricyclic ring systemshaving a total of five to fourteen ring members, wherein at least onering system is aromatic and wherein each ring in the system contains 3to 7 members, such as phenyl, naphthalene, tetrahydronaphthalene andbiphenyl.

By the term “C₅-C₁₂aryl”, used alone or as part of a larger moiety as in“—(CH₂)_(m)C₅-C₁₂aryl”, as used herein, is meant an aromatic groupselected from: phenyl, naphthalene, tehrahydronaphthanlene and biphenyl.

The term “substituted” as used herein, unless otherwise defined, ismeant that the subject chemical moiety has from one to fivesubstituents, suitably from one to three substituents, selected from thegroup consisting of: —CO₂R²⁰, C₁-C₄alkyl, hydroxyC₁-C₄alkyl,C₁-C₄alkyloxy, amino, C₁-C₄alkylamino, aminoC₁-C₄alkyl,diC₁-C₄alkylamino, hydroxy, nitro, tetrazole, cyano, oxo, halogen andtrifluoromethyl, where R²⁰ is selected form hydrogen, C₁-C₄alkyl, andtrifluoromethyl.

Suitably, the term “substituted” as used herein is meant that thesubject chemical moiety has from one to three substituents, selectedfrom the group consisting of: C₁-C₄alkyl, hydroxyC₁-C₄alkyl,C₁-C₄alkyloxy, amino, C₁-C₄alkylamino, aminoC₁-C₄alkyl, hydroxy,tetrazole, halogen and trifluoromethyl.

Suitably, the term “substituted” as used herein is meant that thesubject chemical moiety has one substituent, selected from the groupconsisting of: fluoride and trifluoromethyl.

By the term “heteroatom” as used herein is meant oxygen, nitrogen orsulfur.

By the term “halogen” as used herein is meant a substituent selectedfrom bromide, iodide, chloride and fluoride.

By the term “alkyl” and derivatives thereof and in all carbon chains asused herein, including alkyl chains defined by the term “—(CH₂)_(n)”,“—(CH₂)_(m)” and the like, is meant a linear or branched, saturated orunsaturated hydrocarbon chain, and unless otherwise defined, the carbonchain will contain from 1 to 12 carbon atoms. Examples of alkyl as usedherein include: —CH₃, —CH₂—CH₃, —CH₂—CH₂—CH₃, —CH(CH₃)₂,—CH₂—CH₂—C(CH₃)₃, —C≡C—C(CH₃)₃, —C(CH₃)₃, —(CH₂)₃—CH₃, —CH₂—CH(CH₃)₂,—CH(CH₃)—CH₂—CH₃, —CH═CH₂, and —C≡C—CH₃.

By the term “treating” and derivatives thereof as used herein, is meantprophylatic and therapeutic therapy. Prophylactic therapy isappropriate, for example, when a subject is considered at high risk fordeveloping cancer, or when a subject has been exposed to a carcinogen.

By the phrases “to a therapeutic extent”, “treating” and“therapeutically effective amount” and derivatives thereof as usedherein, unless otherwise defined, is meant that amount of a drug orpharmaceutical agent that will elicit the biological or medical responseof a tissue, system, animal or human that is being sought, for instance,by a researcher or clinician. Furthermore, the term “therapeuticallyeffective amount” means any amount which, as compared to a correspondingsubject who has not received such amount, results in improved treatment,healing, prevention, lessening in severity or amelioration of specifiedcancers.

The pharmaceutically acceptable salts of the compounds of the inventionare readily prepared by those of skill in the art.

Compounds of Formula (I) and Formula (II) are included in thepharmaceutical compositions of the invention and used in the methods ofthe invention. Where a —COOH or —OH group is present, pharmaceuticallyacceptable esters can be employed, for example methyl, ethyl,pivaloyloxymethyl, and the like for —COOH, and acetate maleate and thelike for —OH, and those esters known in the art for modifying solubilityor hydrolysis characteristics, for use as sustained release or prodrugformulations.

The compounds of Formula (I) and Formula (II) are prepared as shown inSchemes 1 to 3 below, or by analogous methods. All of the startingmaterials are commercially available, readily made from commerciallyavailable starting materials by those of skill in the art or preparedaccording to literature reports unless otherwise noted in theexperimental section.

Reagents: (a) Phthalimide, PPh₃, DEAD, THF, RT; (b) NH₂NH₂, MeOH, 50° C.

Amino alcohol (I-1) was reacted under Mitsunobu conditions to providethe differentially protected diamine (I-2). Mitsunobu reactions are wellknow to those skilled in the art of organic synthesis. Methods andreaction conditions for such transformations are discussed in Synthesis1981, 1-28. Selective deprotection of the phthalimide group of (I-2)using a nucleophilic amine such as hydrazine or methyl amine in a polarsolvent such as methanol, afforded amine (I-3). Many differentprotecting groups are available to one skilled in the art and can beused here as long as they do not interfere with the processes listedherein. Methods for the protection of amines are described in standardreference volumes, such as Greene “Protective Groups in OrganicSynthesis” (published by Wiley-Interscience).

Reagents: (a) PyBrop, (i-Pr)₂NEt, 1,1-dimethylethyl(2-amino-3-phenylpropyl)carbamate, DCM, RT; (b)5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole, K₂CO₃,Pd(PPh₃)₄, dioxane/H₂O; (c) HCl/dioxane, MeOH, RT.

Carboxylic acid (II-1) was reacted with 1,1-dimethylethyl(2-amino-3-phenylpropyl)carbamate to form amide (II-2). A variety ofamide coupling reagents such as EDC, PyBrop, etc. are commerciallyavailable. Amide coupling reactions are generally run in solvents suchas DCM or DMF, utilizing an organic base like Et₃N or (i-Pr)₂NEt.Bromide (II-2) was coupled with5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole using aSuzuki coupling procedure. Suzuki-like couplings are typically run usinga palladium(0) catalyst such as Pd(PPh₃)₄ with an inorganic base, forexample K₂CO₃, Na₂CO₃ or K₃PO₄, in an aqueous mixture containingethereal solvents such as DME, dioxane, or THF. Methods forpalladium-mediated couplings are described in standard referencevolumes, such as Schlosser “Organometallics in Synthesis” (published byWiley and sons). Acidic treatment of II-3 with HCl or TFA to remove theBoc protecting group produced amine (II-4). Many different protectinggroups are available to one skilled in the art and can be used here aslong as they do not interfere with the processes listed herein. Methodsfor the protection of amines are described in standard referencevolumes, such as Greene “Protective Groups in Organic Synthesis”(published by Wiley-Interscience).

Reagents: (a) Pd(PPh₃)₂Cl₂, Et₃N, toluene, 100° C.; (b) 2N NaOH, THF;(c) PyBrop, (i-Pr)₂NEt, DCM, RT; (d) NCS, DMF, 50° C.; (e) hydrazinehydrate, MeOH/DCM.

1-Methyl-5-(tributylstannanyl)-1H-1,2,3-triazole (III-I) was coupledwith methyl 4-bromobenzoate (III-2) using a Stille coupling procedure.Stille-like couplings are typically catalyzed by a palladium(0) catalystsuch as Pd(PPh₃)₄ with an organic base, for example Et₃N, in toluene,DMF and dioxane. Methods for palladium-mediated couplings are describedin standard reference volumes, such as Schlosser “Organometallics inSynthesis” (published by Wiley and sons). Carboxylic acid (III-3) wasreacted with2-{(2S)-2-amino-3-[2-(trifluoromethyl)phenyl]propyl}-1H-isoindole-1,3(2H)-dioneto form amide (III-5). A variety of amide coupling reagents such as EDC,PyBrop, etc. are commercially available. Amide coupling reactions aregenerally run in solvents such as DCM or DMF, utilizing an organic baselike Et₃N or (i-Pr)₂NEt. Chlorination of the triazole was realized withNCS in DMF. The treatment of III-6 with hydrazine hydrate to remove thephthilamide protecting group produced amine (III-7).

By the term “co-administering” and derivatives thereof as used herein ismeant either simultaneous administration or any manner of separatesequential administration of an AKT inhibiting compound, as describedherein, and a further active ingredient or ingredients, known to beuseful in the treatment of cancer, including chemotherapy and radiationtreatment, or to be useful in the treatment of arthritis. The termfurther active ingredient or ingredients, as used herein, includes anycompound or therapeutic agent known to or that demonstrates advantageousproperties when administered to a patient in need of treatment forcancer or arthritis. Preferably, if the administration is notsimultaneous, the compounds are administered in a close time proximityto each other. Furthermore, it does not matter if the compounds areadministered in the same dosage form, e.g. one compound may beadministered topically and another compound may be administered orally.

Typically, any anti-neoplastic agent that has activity versus asusceptible tumor being treated may be co-administered in the treatmentof cancer in the present invention. Examples of such agents can be foundin Cancer Principles and Practice of Oncology by V. T. Devita and S.Hellman (editors), 6^(th) edition (Feb. 15, 2001), Lippincott Williams &Wilkins Publishers. A person of ordinary skill in the art would be ableto discern which combinations of agents would be useful based on theparticular characteristics of the drugs and the cancer involved. Typicalanti-neoplastic agents useful in the present invention include, but arenot limited to, anti-microtubule agents such as diterpenoids and vincaalkaloids; platinum coordination complexes; alkylating agents such asnitrogen mustards, oxazaphosphorines, alkylsulfonates, nitrosoureas, andtriazenes; antibiotic agents such as anthracyclins, actinomycins andbleomycins; topoisomerase II inhibitors such as epipodophyllotoxins;antimetabolites such as purine and pyrimidine analogues and anti-folatecompounds; topoisomerase I inhibitors such as camptothecins; hormonesand hormonal analogues; signal transduction pathway inhibitors;non-receptor tyrosine kinase angiogenesis inhibitors; immunotherapeuticagents; proapoptotic agents; and cell cycle signaling inhibitors.

Examples of a further active ingredient or ingredients (anti-neoplasticagent) for use in combination or co-administered with the presentlyinvented AKT inhibiting compounds are chemotherapeutic agents.

Anti-microtubule or anti-mitotic agents are phase specific agents activeagainst the microtubules of tumor cells during M or the mitosis phase ofthe cell cycle. Examples of anti-microtubule agents include, but are notlimited to, diterpenoids and vinca alkaloids.

Diterpenoids, which are derived from natural sources, are phase specificanti-cancer agents that operate at the G₂/M phases of the cell cycle. Itis believed that the diterpenoids stabilize the β-tubulin subunit of themicrotubules, by binding with this protein. Disassembly of the proteinappears then to be inhibited with mitosis being arrested and cell deathfollowing. Examples of diterpenoids include, but are not limited to,paclitaxel and its analog docetaxel.

Paclitaxel, 5β,20-epoxy-1,2α,4,7β,10β,13α-hexa-hydroxytax-11-en-9-one4,10-diacetate 2-benzoate 13-ester with(2R,3S)-N-benzoyl-3-phenylisoserine; is a natural diterpene productisolated from the Pacific yew tree Taxus brevifolia and is commerciallyavailable as an injectable solution TAXOL®. It is a member of the taxanefamily of terpenes. It was first isolated in 1971 by Wani et al. J. Am.Chem, Soc., 93:2325. 1971), who characterized its structure by chemicaland X-ray crystallographic methods. One mechanism for its activityrelates to paclitaxel's capacity to bind tubulin, thereby inhibitingcancer cell growth. Schiff et al., Proc. Natl, Acad, Sci. USA,77:1561-1565 (1980); Schiff et al., Nature, 277:665-667 (1979); Kumar,J. Biol, Chem, 256: 10435-10441 (1981). For a review of synthesis andanticancer activity of some paclitaxel derivatives see: D. G. I.Kingston et al., Studies in Organic Chemistry vol. 26, entitled “Newtrends in Natural Products Chemistry 1986”, Attaur-Rahman, P. W. LeQuesne, Eds. (Elsevier, Amsterdam, 1986) pp 219-235.

Paclitaxel has been approved for clinical use in the treatment ofrefractory ovarian cancer in the United States (Markman et al., YaleJournal of Biology and Medicine, 64:583, 1991; McGuire et al., Ann.Intern, Med., 111:273, 1989) and for the treatment of breast cancer(Holmes et al., J. Nat. Cancer Inst., 83:1797, 1991.) It is a potentialcandidate for treatment of neoplasms in the skin (Einzig et. al., Proc.Am. Soc. Clin. Oncol., 20:46) and head and neck carcinomas (Forastireet. al., Sem. Oncol., 20:56, 1990). The compound also shows potentialfor the treatment of polycystic kidney disease (Woo et. al., Nature,368:750. 1994), lung cancer and malaria. Treatment of patients withpaclitaxel results in bone marrow suppression (multiple cell lineages,Ignoff, R. J. et. al, Cancer Chemotherapy Pocket Guide. 1998) related tothe duration of dosing above a threshold concentration (50 nM) (Kearns,C. M. et. al., Seminars in Oncology, 3(6) p. 16-23, 1995).

Docetaxel, (2R,3S)-N-carboxy-3-phenylisoserine,N-tert-butyl ester,13-ester with 5β-20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytax-11-en-9-one4-acetate 2-benzoate, trihydrate; is commercially available as aninjectable solution as TAXOTERE®. Docetaxel is indicated for thetreatment of breast cancer. Docetaxel is a semisynthetic derivative ofpaclitaxel q.v., prepared using a natural precursor,10-deacetyl-baccatin III, extracted from the needle of the European Yewtree. The dose limiting toxicity of docetaxel is neutropenia.

Vinca alkaloids are phase specific anti-neoplastic agents derived fromthe periwinkle plant. Vinca alkaloids act at the M phase (mitosis) ofthe cell cycle by binding specifically to tubulin. Consequently, thebound tubulin molecule is unable to polymerize into microtubules.Mitosis is believed to be arrested in metaphase with cell deathfollowing. Examples of vinca alkaloids include, but are not limited to,vinblastine, vincristine, and vinorelbine.

Vinblastine, vincaleukoblastine sulfate, is commercially available asVELBAN® as an injectable solution. Although, it has possible indicationas a second line therapy of various solid tumors, it is primarilyindicated in the treatment of testicular cancer and various lymphomasincluding Hodgkin's Disease; and lymphocytic and histiocytic lymphomas.Myelosuppression is the dose limiting side effect of vinblastine.

Vincristine, vincaleukoblastine, 22-oxo-, sulfate, is commerciallyavailable as ONCOVIN® as an injectable solution. Vincristine isindicated for the treatment of acute leukemias and has also found use intreatment regimens for Hodgkin's and non-Hodgkin's malignant lymphomas.Alopecia and neurologic effects are the most common side effect ofvincristine and to a lesser extent myelosupression and gastrointestinalmucositis effects occur.

Vinorelbine,3′,4′-didehydro-4′-deoxy-C′-norvincaleukoblastine[R-(R*,R*)-2,3-dihydroxybutanedioate(1:2)(salt)], commercially available as an injectable solution ofvinorelbine tartrate (NAVELBINE®), is a semisynthetic vinca alkaloid.Vinorelbine is indicated as a single agent or in combination with otherchemotherapeutic agents, such as cisplatin, in the treatment of varioussolid tumors, particularly non-small cell lung, advanced breast, andhormone refractory prostate cancers. Myelosuppression is the most commondose limiting side effect of vinorelbine.

Platinum coordination complexes are non-phase specific anti-canceragents, which are interactive with DNA. The platinum complexes entertumor cells, undergo, aquation and form intra- and interstrandcrosslinks with DNA causing adverse biological effects to the tumor.Examples of platinum coordination complexes include, but are not limitedto, cisplatin and carboplatin.

Cisplatin, cis-diamminedichloroplatinum, is commercially available asPLATINOL® as an injectable solution. Cisplatin is primarily indicated inthe treatment of metastatic testicular and ovarian cancer and advancedbladder cancer. The primary dose limiting side effects of cisplatin arenephrotoxicity, which may be controlled by hydration and diuresis, andototoxicity.

Carboplatin, platinum, diammine [1,1-cyclobutane-dicarboxylate(2)-O,O′],is commercially available as PARAPLATIN® as an injectable solution.Carboplatin is primarily indicated in the first and second linetreatment of advanced ovarian carcinoma. Bone marrow suppression is thedose limiting toxicity of carboplatin.

Alkylating agents are non-phase anti-cancer specific agents and strongelectrophiles. Typically, alkylating agents form covalent linkages, byalkylation, to DNA through nucleophilic moieties of the DNA moleculesuch as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazolegroups. Such alkylation disrupts nucleic acid function leading to celldeath. Examples of alkylating agents include, but are not limited to,nitrogen mustards such as cyclophosphamide, melphalan, and chlorambucil;alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; andtriazenes such as dacarbazine.

Cyclophosphamide,2-[bis(2-chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine 2-oxidemonohydrate, is commercially available as an injectable solution ortablets as CYTOXAN®. Cyclophosphamide is indicated as a single agent orin combination with other chemotherapeutic agents, in the treatment ofmalignant lymphomas, multiple myeloma, and leukemias. Alopecia, nausea,vomiting and leukopenia are the most common dose limiting side effectsof cyclophosphamide.

Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commerciallyavailable as an injectable solution or tablets as ALKERAN®. Melphalan isindicated for the palliative treatment of multiple myeloma andnon-resectable epithelial carcinoma of the ovary. Bone marrowsuppression is the most common dose limiting side effect of melphalan.

Chlorambucil, 4-[bis(2-chloroethyl)amino]benzenebutanoic acid, iscommercially available as LEUKERAN® tablets. Chlorambucil is indicatedfor the palliative treatment of chronic lymphatic leukemia, andmalignant lymphomas such as lymphosarcoma, giant follicular lymphoma,and Hodgkin's disease. Bone marrow suppression is the most common doselimiting side effect of chlorambucil.

Busulfan, 1,4-butanediol dimethanesulfonate, is commercially availableas MYLERAN® TABLETS. Busulfan is indicated for the palliative treatmentof chronic myelogenous leukemia. Bone marrow suppression is the mostcommon dose limiting side effects of busulfan.

Carmustine, 1,3-[bis(2-chloroethyl)-1-nitrosourea, is commerciallyavailable as single vials of lyophilized material as BiCNU®. Carmustineis indicated for the palliative treatment as a single agent or incombination with other agents for brain tumors, multiple myeloma,Hodgkin's disease, and non-Hodgkin's lymphomas. Delayed myelosuppressionis the most common dose limiting side effects of carmustine.

Dacarbazine, 5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide, iscommercially available as single vials of material as DTIC-Dome®.Dacarbazine is indicated for the treatment of metastatic malignantmelanoma and in combination with other agents for the second linetreatment of Hodgkin's Disease. Nausea, vomiting, and anorexia are themost common dose limiting side effects of dacarbazine.

Antibiotic anti-neoplastics are non-phase specific agents, which bind orintercalate with DNA. Typically, such action results in stable DNAcomplexes or strand breakage, which disrupts ordinary function of thenucleic acids leading to cell death. Examples of antibioticanti-neoplastic agents include, but are not limited to, actinomycinssuch as dactinomycin, anthrocyclins such as daunorubicin anddoxorubicin; and bleomycins.

Dactinomycin, also know as Actinomycin D, is commercially available ininjectable form as COSMEGEN®. Dactinomycin is indicated for thetreatment of Wilm's tumor and rhabdomyosarcoma. Nausea, vomiting, andanorexia are the most common dose limiting side effects of dactinomycin.

Daunorubicin,(8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12naphthacenedione hydrochloride, is commercially available as a liposomalinjectable form as DAUNOXOME® or as an injectable as CERUBIDINE®.Daunorubicin is indicated for remission induction in the treatment ofacute nonlymphocytic leukemia and advanced HIV associated Kaposi'ssarcoma. Myelosuppression is the most common dose limiting side effectof daunorubicin.

Doxorubicin,(8S,10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-8-glycoloyl,7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12 naphthacenedionehydrochloride, is commercially available as an injectable form as RUBEX®or ADRIAMYCIN RDF®. Doxorubicin is primarily indicated for the treatmentof acute lymphoblastic leukemia and acute myeloblastic leukemia, but isalso a useful component in the treatment of some solid tumors andlymphomas. Myelosuppression is the most common dose limiting side effectof doxorubicin.

Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated froma strain of Streptomyces verticillus, is commercially available asBLENOXANE®. Bleomycin is indicated as a palliative treatment, as asingle agent or in combination with other agents, of squamous cellcarcinoma, lymphomas, and testicular carcinomas. Pulmonary and cutaneoustoxicities are the most common dose limiting side effects of bleomycin.

Topoisomerase II inhibitors include, but are not limited to,epipodophyllotoxins.

Epipodophyllotoxins are phase specific anti-neoplastic agents derivedfrom the mandrake plant. Epipodophyllotoxins typically affect cells inthe S and G₂ phases of the cell cycle by forming a ternary complex withtopoisomerase II and DNA causing DNA strand breaks. The strand breaksaccumulate and cell death follows. Examples of epipodophyllotoxinsinclude, but are not limited to, etoposide and teniposide.

Etoposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-ethylidene-β-D-glucopyranoside], is commercially availableas an injectable solution or capsules as VePESID® and is commonly knownas VP-16. Etoposide is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of testicular andnon-small cell lung cancers. Myelosuppression is the most common sideeffect of etoposide. The incidence of leucopenia tends to be more severethan thrombocytopenia.

Teniposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-thenylidene-β-D-glucopyranoside], is commercially availableas an injectable solution as VUMON® and is commonly known as VM-26.Teniposide is indicated as a single agent or in combination with otherchemotherapy agents in the treatment of acute leukemia in children.Myelosuppression is the most common dose limiting side effect ofteniposide. Teniposide can induce both leucopenia and thrombocytopenia.

Antimetabolite neoplastic agents are phase specific anti-neoplasticagents that act at S phase (DNA synthesis) of the cell cycle byinhibiting DNA synthesis or by inhibiting purine or pyrimidine basesynthesis and thereby limiting DNA synthesis. Consequently, S phase doesnot proceed and cell death follows. Examples of antimetaboliteanti-neoplastic agents include, but are not limited to, fluorouracil,methotrexate, cytarabine, mecaptopurine, thioguanine, and gemcitabine.

5-fluorouracil, 5-fluoro-2,4-(1H,3H) pyrimidinedione, is commerciallyavailable as fluorouracil. Administration of 5-fluorouracil leads toinhibition of thymidylate synthesis and is also incorporated into bothRNA and DNA. The result typically is cell death. 5-fluorouracil isindicated as a single agent or in combination with other chemotherapyagents in the treatment of carcinomas of the breast, colon, rectum,stomach and pancreas. Myelosuppression and mucositis are dose limitingside effects of 5-fluorouracil. Other fluoropyrimidine analogs include5-fluoro deoxyuridine (floxuridine) and 5-fluorodeoxyuridinemonophosphate.

Cytarabine, 4-amino-1-β-D-arabinofuranosyl-2 (1H)-pyrimidinone, iscommercially available as CYTOSAR-U® and is commonly known as Ara-C. Itis believed that cytarabine exhibits cell phase specificity at S-phaseby inhibiting DNA chain elongation by terminal incorporation ofcytarabine into the growing DNA chain. Cytarabine is indicated as asingle agent or in combination with other chemotherapy agents in thetreatment of acute leukemia. Other cytidine analogs include5-azacytidine and 2′,2′-difluorodeoxycytidine (gemcitabine). Cytarabineinduces leucopenia, thrombocytopenia, and mucositis.

Mercaptopurine, 1,7-dihydro-6H-purine-6-thione monohydrate, iscommercially available as PURINETHOL®. Mercaptopurine exhibits cellphase specificity at S-phase by inhibiting DNA synthesis by an as of yetunspecified mechanism. Mercaptopurine is indicated as a single agent orin combination with other chemotherapy agents in the treatment of acuteleukemia. Myelosuppression and gastrointestinal mucositis are expectedside effects of mercaptopurine at high doses. A useful mercaptopurineanalog is azathioprine.

Thioguanine, 2-amino-1,7-dihydro-6H-purine-6-thione, is commerciallyavailable as TABLOID®. Thioguanine exhibits cell phase specificity atS-phase by inhibiting DNA synthesis by an as of yet unspecifiedmechanism. Thioguanine is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of acute leukemia.Myelosuppression, including leucopenia, thrombocytopenia, and anemia, isthe most common dose limiting side effect of thioguanine administration.However, gastrointestinal side effects occur and can be dose limiting.Other purine analogs include pentostatin, erythrohydroxynonyladenine,fludarabine phosphate, and cladribine.

Gemcitabine, 2′-deoxy-2′,2′-difluorocytidine monohydrochloride(β-isomer), is commercially available as GEMZAR®. Gemcitabine exhibitscell phase specificity at S-phase and by blocking progression of cellsthrough the G1/S boundary. Gemcitabine is indicated in combination withcisplatin in the treatment of locally advanced non-small cell lungcancer and alone in the treatment of locally advanced pancreatic cancer.Myelosuppression, including leucopenia, thrombocytopenia, and anemia, isthe most common dose limiting side effect of gemcitabine administration.

Methotrexate,N-[4[[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamicacid, is commercially available as methotrexate sodium. Methotrexateexhibits cell phase effects specifically at S-phase by inhibiting DNAsynthesis, repair and/or replication through the inhibition ofdyhydrofolic acid reductase which is required for synthesis of purinenucleotides and thymidylate. Methotrexate is indicated as a single agentor in combination with other chemotherapy agents in the treatment ofchoriocarcinoma, meningeal leukemia, non-Hodgkin's lymphoma, andcarcinomas of the breast, head, neck, ovary and bladder.Myelosuppression (leucopenia, thrombocytopenia, and anemia) andmucositis are expected side effect of methotrexate administration.

Camptothecins, including, camptothecin and camptothecin derivatives areavailable or under development as Topoisomerase I inhibitors.Camptothecins cytotoxic activity is believed to be related to itsTopoisomerase I inhibitory activity. Examples of camptothecins include,but are not limited to irinotecan, topotecan, and the various opticalforms of7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20-camptothecindescribed below.

Irinotecan HCl,(4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)carbonyloxy]-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dionehydrochloride, is commercially available as the injectable solutionCAMPTOSAR®.

Irinotecan is a derivative of camptothecin which binds, along with itsactive metabolite SN-38, to the topoisomerase I-DNA complex. It isbelieved that cytotoxicity occurs as a result of irreparable doublestrand breaks caused by interaction of the topoisomerase I:DNA:irintecanor SN-38 ternary complex with replication enzymes. Irinotecan isindicated for treatment of metastatic cancer of the colon or rectum. Thedose limiting side effects of irinotecan HCl are myelosuppression,including neutropenia, and GI effects, including diarrhea.

Topotecan HCl,(S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14-(4H,12H)-dionemonohydrochloride, is commercially available as the injectable solutionHYCAMTIN®. Topotecan is a derivative of camptothecin which binds to thetopoisomerase I-DNA complex and prevents religation of singles strandbreaks caused by Topoisomerase I in response to torsional strain of theDNA molecule. Topotecan is indicated for second line treatment ofmetastatic carcinoma of the ovary and small cell lung cancer. The doselimiting side effect of topotecan HCl is myelosuppression, primarilyneutropenia.

Also of interest, is the camptothecin derivative of formula A following,currently under development, including the racemic mixture (R,S) form aswell as the R and S enantiomers:

known by the chemical name“7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(R,S)-camptothecin(racemic mixture) or“7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(R)-camptothecin(R enantiomer) or“7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(S)-camptothecin(S enantiomer). Such compound as well as related compounds aredescribed, including methods of making, in U.S. Pat. Nos. 6,063,923;5,342,947; 5,559,235; 5,491,237 and pending U.S. patent Application No.Ser. 08/977,217 filed Nov. 24, 1997.

Hormones and hormonal analogues are useful compounds for treatingcancers in which there is a relationship between the hormone(s) andgrowth and/or lack of growth of the cancer. Examples of hormones andhormonal analogues useful in cancer treatment include, but are notlimited to, adrenocorticosteroids such as prednisone and prednisolonewhich are useful in the treatment of malignant lymphoma and acuteleukemia in children; aminoglutethimide and other aromatase inhibitorssuch as anastrozole, letrazole, vorazole, and exemestane useful in thetreatment of adrenocortical carcinoma and hormone dependent breastcarcinoma containing estrogen receptors; progestrins such as megestrolacetate useful in the treatment of hormone dependent breast cancer andendometrial carcinoma; estrogens, androgens, and anti-androgens such asflutamide, nilutamide, bicalutamide, cyproterone acetate and5α-reductases such as finasteride and dutasteride, useful in thetreatment of prostatic carcinoma and benign prostatic hypertrophy;anti-estrogens such as tamoxifen, toremifene, raloxifene, droloxifene,iodoxyfene, as well as selective estrogen receptor modulators (SERMS)such those described in U.S. Pat. Nos. 5,681,835, 5,877,219, and6,207,716, useful in the treatment of hormone dependent breast carcinomaand other susceptible cancers; and gonadotropin-releasing hormone (GnRH)and analogues thereof which stimulate the release of leutinizing hormone(LH) and/or follicle stimulating hormone (FSH) for the treatmentprostatic carcinoma, for instance, LHRH agonists and antagagonists suchas goserelin acetate and luprolide.

Signal transduction pathway inhibitors are those inhibitors, which blockor inhibit a chemical process which evokes an intracellular change. Asused herein this change is cell proliferation or differentiation. Signaltranduction inhibitors useful in the present invention includeinhibitors of receptor tyrosine kinases, non-receptor tyrosine kinases,SH2/SH3domain blockers, serine/threonine kinases, phosphotidylinositol-3 kinases, myo-inositol signaling, and Ras oncogenes.

Several protein tyrosine kinases catalyse the phosphorylation ofspecific tyrosyl residues in various proteins involved in the regulationof cell growth. Such protein tyrosine kinases can be broadly classifiedas receptor or non-receptor kinases.

Receptor tyrosine kinases are transmembrane proteins having anextracellular ligand binding domain, a transmembrane domain, and atyrosine kinase domain. Receptor tyrosine kinases are involved in theregulation of cell growth and are generally termed growth factorreceptors. Inappropriate or uncontrolled activation of many of thesekinases, i.e. aberrant kinase growth factor receptor activity, forexample by over-expression or mutation, has been shown to result inuncontrolled cell growth. Accordingly, the aberrant activity of suchkinases has been linked to malignant tissue growth. Consequently,inhibitors of such kinases could provide cancer treatment methods.Growth factor receptors include, for example, epidermal growth factorreceptor (EGFr), platelet derived growth factor receptor (PDGFr), erbB2,erbB4, vascular endothelial growth factor receptor (VEGFr), tyrosinekinase with immunoglobulin-like and epidermal growth factor homologydomains (TIE-2), insulin growth factor-I (IGFI) receptor, macrophagecolony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growthfactor (FGF) receptors, Trk receptors (TrkA, TrkB, and TrkC), ephrin(eph) receptors, and the RET protooncogene. Several inhibitors of growthreceptors are under development and include ligand antagonists,antibodies, tyrosine kinase inhibitors and anti-sense oligonucleotides.Growth factor receptors and agents that inhibit growth factor receptorfunction are described, for instance, in Kath, John C., Exp. Opin. Ther.Patents (2000) 10(6):803-818; Shawver et al DDT Vol 2, No. 2 February1997; and Lofts, F. J. et al, “Growth factor receptors as targets”, NewMolecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr,David, CRC press 1994, London.

Tyrosine kinases, which are not growth factor receptor kinases aretermed non-receptor tyrosine kinases. Non-receptor tyrosine kinases foruse in the present invention, which are targets or potential targets ofanti-cancer drugs, include cSrc, Lck, Fyn, Yes, Jak, cAbl, FAK (Focaladhesion kinase), Brutons tyrosine kinase, and Bcr-Abl. Suchnon-receptor kinases and agents which inhibit non-receptor tyrosinekinase function are described in Sinh, S. and Corey, S. J., (1999)Journal of Hematotherapy and Stem Cell Research 8 (5): 465-80; andBolen, J. B., Brugge, J. S., (1997) Annual review of Immunology. 15:371-404.

SH2/SH3 domain blockers are agents that disrupt SH2 or SH3 domainbinding in a variety of enzymes or adaptor proteins including, PI3-K p85subunit, Src family kinases, adaptor molecules (Shc, Crk, Nck, Grb2) andRas-GAP. SH2/SH3 domains as targets for anti-cancer drugs are discussedin Smithgall, T. E. (1995), Journal of Pharmacological and ToxicologicalMethods. 34(3) 125-32.

Inhibitors of Serine/Threonine Kinases including MAP kinase cascadeblockers which include blockers of Raf kinases (rafk), Mitogen orExtracellular Regulated Kinase (MEKs), and Extracellular RegulatedKinases (ERKs); and Protein kinase C family member blockers includingblockers of PKCs (alpha, beta, gamma, epsilon, mu, lambda, iota, zeta).IkB kinase family (IKKa, IKKb), PKB family kinases, akt kinase familymembers, and TGF beta receptor kinases. Such Serine/Threonine kinasesand inhibitors thereof are described in Yamamoto, T., Taya, S.,Kaibuchi, K., (1999), Journal of Biochemistry. 126 (5) 799-803; Brodt,P, Samani, A., and Navab, R. (2000), Biochemical Pharmacology, 60.1101-1107; Massague, J., Weis-Garcia, F. (1996) Cancer Surveys.27:41-64; Philip, P. A., and Harris, A. L. (1995), Cancer Treatment andResearch. 78: 3-27, Lackey, K. et al Bioorganic and Medicinal ChemistryLetters, (10), 2000, 223-226; U.S. Pat. No. 6,268,391; andMartinez-lacaci, L., et al, Int. J. Cancer (2000), 88(1), 44-52.

Inhibitors of Phosphotidyl inositol-3 Kinase family members includingblockers of PI3-kinase, ATM, DNA-PK, and Ku may also be useful in thepresent invention. Such kinases are discussed in Abraham, R. T. (1996),Current Opinion in Immunology. 8 (3) 412-8; Canman, C. E., Lim, D. S.(1998), Oncogene 17 (25) 3301-3308; Jackson, S. P. (1997), InternationalJournal of Biochemistry and Cell Biology. 29 (7):935-8; and Zhong, H. etal, Cancer res, (2000) 60(6), 1541-1545.

Also of interest in the present invention are Myo-inositol signalinginhibitors such as phospholipase C blockers and Myoinositol analogues.Such signal inhibitors are described in Powis, G., and Kozikowski A.,(1994) New Molecular Targets for Cancer Chemotherapy ed., Paul Workmanand David Kerr, CRC press 1994, London.

Another group of signal transduction pathway inhibitors are inhibitorsof Ras Oncogene. Such inhibitors include inhibitors offarnesyltransferase, geranyl-geranyl transferase, and CAAX proteases aswell as anti-sense oligonucleotides, ribozymes and immunotherapy. Suchinhibitors have been shown to block ras activation in cells containingwild type mutant ras, thereby acting as antiproliferation agents. Rasoncogene inhibition is discussed in Scharovsky, O. G., Rozados, V. R.,Gervasoni, S. I. Matar, P. (2000), Journal of Biomedical Science. 7(4)292-8; Ashby, M. N. (1998), Current Opinion in Lipidology. 9 (2) 99-102;and BioChim. Biophys. Acta, (19899) 1423(3):19-30.

As mentioned above, antibody antagonists to receptor kinase ligandbinding may also serve as signal transduction inhibitors. This group ofsignal transduction pathway inhibitors includes the use of humanizedantibodies to the extracellular ligand binding domain of receptortyrosine kinases. For example Imclone C225 EGFR specific antibody (seeGreen, M. C. et al, Monoclonal Antibody Therapy for Solid Tumors, CancerTreat. Rev., (2000), 26(4), 269-286); Herceptin® erbB2 antibody (seeTyrosine Kinase Signalling in Breast cancer:erbB Family ReceptorTyrosine Kniases, Breast cancer Res., 2000, 2(3), 176-183); and 2CBVEGFR2 specific antibody (see Brekken, R. A. et al, Selective Inhibitionof VEGFR2 Activity by a monoclonal Anti-VEGF antibody blocks tumorgrowth in mice, Cancer Res. (2000) 60, 5117-5124).

Non-receptor kinase angiogenesis inhibitors may also be useful in thepresent invention. Inhibitors of angiogenesis related VEGFR and TIE2 arediscussed above in regard to signal transduction inhibitors (bothreceptors are receptor tyrosine kinases). Angiogenesis in general islinked to erbB2/EGFR signaling since inhibitors of erbB2 and EGFR havebeen shown to inhibit angiogenesis, primarily VEGF expression.Accordingly, non-receptor tyrosine kinase inhibitors may be used incombination with the compounds of the present invention. For example,anti-VEGF antibodies, which do not recognize VEGFR (the receptortyrosine kinase), but bind to the ligand; small molecule inhibitors ofintegrin (alpha_(v) beta₃) that will inhibit angiogenesis; endostatinand angiostatin (non-RTK) are also useful in combination with thecompounds disclosed herein. (See Bruns C J et al (2000), Cancer Res.,60: 2926-2935; Schreiber A B, Winkler M E, and Derynck R. (1986),Science, 232: 1250-1253; Yen L et al. (2000), Oncogene 19: 3460-3469).

Agents used in immunotherapeutic regimens may also be useful incombination with the compounds of Formula (I) and Formula (II). Thereare a number of immunologic strategies to generate an immune response.These strategies are generally in the realm of tumor vaccinations. Theefficacy of immunologic approaches may be greatly enhanced throughcombined inhibition of signaling pathways using a small moleculeinhibitor. Discussion of the immunologic/tumor vaccine approach againsterbB2/EGFR are found in Reilly R T et al. (2000), Cancer Res. 60:3569-3576; and Chen Y, Hu D, Eling D J, Robbins J, and Kipps T J.(1998), Cancer Res. 58: 1965-1971.

Agents used in proapoptotic regimens (e.g., bcl-2 antisenseoligonucleotides) may also be used in the combination of the presentinvention. Members of the Bcl-2 family of proteins block apoptosis.Upregulation of bcl-2 has therefore been linked to chemoresistance.Studies have shown that the epidermal growth factor (EGF) stimulatesanti-apoptotic members of the bcl-2 family (i.e., mcl-1). Therefore,strategies designed to downregulate the expression of bcl-2 in tumorshave demonstrated clinical benefit and are now in Phase II/III trials,namely Genta's G3139 bcl-2 antisense oligonucleotide. Such proapoptoticstrategies using the antisense oligonucleotide strategy for bcl-2 arediscussed in Water J S et al. (2000), J. Clin. Oncol. 18: 1812-1823; andKitada S et al. (1994), Antisense Res. Dev. 4: 71-79.

Cell cycle signalling inhibitors inhibit molecules involved in thecontrol of the cell cycle. A family of protein kinases called cyclindependent kinases (CDKs) and their interaction with a family of proteinstermed cyclins controls progression through the eukaryotic cell cycle.The coordinate activation and inactivation of different cyclin/CDKcomplexes is necessary for normal progression through the cell cycle.Several inhibitors of cell cycle signalling are under development. Forinstance, examples of cyclin dependent kinases, including CDK2, CDK4,and CDK6 and inhibitors for the same are described in, for instance,Rosania et al, Exp. Opin. Ther. Patents (2000) 10(2):215-230.

In one embodiment, the cancer treatment method of the claimed inventionincludes the co-administration a compound of Formula (I) or Formula (II)and/or a pharmaceutically acceptable salt, hydrate, solvate or pro-drugthereof and at least one anti-neoplastic agent, such as one selectedfrom the group consisting of anti-microtubule agents, platinumcoordination complexes, alkylating agents, antibiotic agents,topoisomerase II inhibitors, antimetabolites, topoisomerase Iinhibitors, hormones and hormonal analogues, signal transduction pathwayinhibitors, non-receptor tyrosine kinase angiogenesis inhibitors,immunotherapeutic agents, proapoptotic agents, and cell cycle signalinginhibitors.

Because the pharmaceutically active compounds of the present inventionare active as AKT inhibitors they exhibit therapeutic utility intreating cancer and arthritis.

The present invention therefore provides a method of treating cancer ina mammal, including a human, including wherein the cancer is selectedfrom: brain (gliomas), glioblastomas, leukemias, Bannayan-Zonanasyndrome, Cowden disease, Lhermitte-Duclos disease, breast, inflammatorybreast cancer, Wilm's tumor, Ewing's sarcoma, Rhabdomyosarcoma,ependymoma, medulloblastoma, colon, head and neck, kidney, lung, liver,melanoma, ovarian, pancreatic, prostate, sarcoma, osteosarcoma, giantcell tumor of bone, thyroid,

Lymphoblastic T cell leukemia, Chronic myelogenous leukemia, Chroniclymphocytic leukemia, Hairy-cell leukemia, acute lymphoblastic leukemia,acute myelogenous leukemia, Chronic neutrophilic leukemia, Acutelymphoblastic T cell leukemia, Plasmacytoma, Immunoblastic large cellleukemia, Mantle cell leukemia, Multiple myeloma Megakaryoblasticleukemia, multiple myeloma, Acute megakaryocytic leukemia, promyelocyticleukemia, Erythroleukemia,

malignant lymphoma, hodgkins lymphoma, non-hodgkins lymphoma,lymphoblastic T cell lymphoma, Burkitt's lymphoma, follicular lymphoma,

neuroblastoma, bladder cancer, urothelial cancer, lung cancer, vulvalcancer, cervical cancer, endometrial cancer, renal cancer, mesothelioma,esophageal cancer, salivary gland cancer, hepatocellular cancer, gastriccancer, nasopharangeal cancer, buccal cancer, cancer of the mouth, GIST(gastrointestinal stromal tumor) and testicular cancer,

which comprises the administration an effective amount of a presentlyinvented AKT inhibiting compound.

Suitably, the present invention relates to a method for treating acancer selected from brain (gliomas), glioblastomas, leukemias,Bannayan-Zonana syndrome, Cowden disease, Lhermitte-Duclos disease,breast, colon, head and neck, kidney, lung, liver, melanoma, ovarian,pancreatic, prostate, sarcoma and thyroid.

Suitably, the present invention relates to a method for treating acancer selected from breast, ovarian, pancreatic and prostate.

Isolation and Purification of His-Tagged AKT1 (aa 136-480)

Insect cells expressing His-tagged AKT1 (aa 136-480) were lysed in 25 mMHEPES, 100 mM NaCl, 20 mM imidazole; pH 7.5 using a polytron (5 mLslysis buffer/g cells). Cell debris was removed by centrifuging at28,000×g for 30 minutes. The supernatant was filtered through a4.5-micron filter then loaded onto a nickel-chelating columnpre-equilibrated with lysis buffer. The column was washed with 5 columnvolumes (CV) of lysis buffer then with 5 CV of 20% buffer B, wherebuffer B is 25 mM HEPES, 100 mM NaCl, 300 mM imidazole; pH 7.5.His-tagged AKT1 (aa 136-480) was eluted with a 20-100% linear gradientof buffer B over 10 CV. His-tagged AKT1 (136-480) eluting fractions werepooled and diluted 3-fold with buffer C, where buffer C is 25 mM HEPES,pH 7.5. The sample was then chromatographed over a Q-Sepharose HP columnpre-equilibrated with buffer C. The column was washed with 5 CV ofbuffer C then step eluted with 5 CV 10% D, 5 CV 20% D, 5 CV 30% D, 5 CV50% D and 5 CV of 100% D; where buffer D is 25 mM HEPES, 1000 mM NaCl;pH 7.5. His-tagged AKT1 (aa 136-480) containing fractions were pooledand concentrated in a 10-kDa molecular weight cutoff concentrator.His-tagged AKT1 (aa 136-480) was chromatographed over a Superdex 75 gelfiltration column pre-equilibrated with 25 mM HEPES, 200 mM NaCl, 1 mMDTT; pH 7.5. His-tagged AKT1 (aa 136-480) fractions were examined usingSDS-PAGE and mass spec. The protein was pooled, concentrated and frozenat −80 C.

His-tagged AKT2 (aa 138-481) and His-tagged AKT3 (aa 135-479) wereisolated and purified in a similar fashion.

His-Tagged AKT Enzyme Assay

Compounds of the present invention were tested for AKT 1, 2, and 3protein serine kinase inhibitory activity in substrate phosphorylationassays. This assay examines the ability of small molecule organiccompounds to inhibit the serine phosphorylation of a peptide substrate.The substrate phosphorylation assays use the catalytic domains of AKT 1,2, or 3. AKT 1, 2 and 3 are also commercially available from UpstateUSA, Inc. The method measures the ability of the isolated enzyme tocatalyze the transfer of the gamma-phosphate from ATP onto the serineresidue of a biotinylated synthetic peptide SEQ. ID NO: 1(Biotin-ahx-ARKRERAYSFGHHA-amide). Substrate phosphorylation wasdetected by the following procedure:

Assays were performed in 384well U-bottom white plates. 10 nM activatedAKT enzyme was incubated for 40 minutes at room temperature in an assayvolume of 20 ul containing 50 mM MOPS, pH 7.5, 20 mM MgCl₂, 4 uM ATP, 8uM peptide, 0.04 uCi [g-³³P] ATP/well, 1 mM CHAPS, 2 mM DTT, and 1 ul oftest compound in 100% DMSO. The reaction was stopped by the addition of50 ul SPA bead mix (Dulbecco's PBS without Mg²⁺ and Ca²⁺, 0.1% TritonX-100, 5 mM EDTA, 50 uM ATP, 2.5 mg/ml Streptavidin-coated SPA beads.)The plate was sealed, the beads were allowed to settle overnight, andthen the plate was counted in a Packard Topcount MicroplateScintillation Counter (Packard Instrument Co., Meriden, Conn.).

The data for dose responses were plotted as % Control calculated withthe data reduction formula 100*(U1−C2)/(C1−C2) versus concentration ofcompound where U is the unknown value, C1 is the average control valueobtained for DMSO, and C2 is the average control value obtained for 0.1MEDTA. Data are fitted to the curve described by: y=((Vmax*x)/(K+x))where Vmax is the upper asymptote and K is the IC50.

Cloning of Full-Length Human (FL) AKT1:

Full-length human AKT1 gene was amplified by PCR from a plasmidcontaining myristylated-AKT1-ER (gift from Robert T. Abraham, DukeUniversity under MTA, described in Klippel et al. in Molecular andCellular Biology 1998 Volume 18 p. 5699) using the 5′ primer: SEQ. IDNO: 2 5′ TATATAGGATCCATGAGCGACGTGGC 3′ and the 3′ primer: SEQ. ID NO: 3AAATTTCTCGAGTCAGGCCGTGCTGCTGG 3′. The 5′ primer included a BamHI siteand the 3′primer included an XhoI site for cloning purposes. Theresultant PCR product was subcloned in pcDNA3 as a BamHI/XhoI fragment.A mutation in the sequence (TGC) coding for a Cysteine²⁵ was convertedto the wild-type AKT1 sequence (CGC) coding for an Arginine²⁵ bysite-directed mutagenesis using the QuikChange® Site DirectedMutagenesis Kit (Stratagene). The AKT1 mutagenic primer: SEQ. ID NO: 45′ ACCTGGCGGCCACGCTACTTCCTCC and selection primer: SEQ. ID NO: 5 5′CTCGAGCATGCAACTAGAGGGCC (designed to destroy an XbaI site in themultiple cloning site of pcDNA3) were used according to manufacturer'ssuggestions. For expression/purification purposes, AKT1 was isolated asa BamHI/XhoI fragment and cloned into the BamHI/XhoI sites ofpFastbacHTb (Invitrogen).

Expression of FL Human AKT1:

Expression was done using the BAC-to-BAC Baculovirus Expression Systemfrom Invitrogen (catalog #10359-016). Briefly 1) the cDNA wastransferred from the FastBac vector into bacmid DNA, 2) the bacmid DNAwas isolated and used to transfect Sf9 insect cells, 3) the virus wasproduced in Sf9 cells, 4) T. ni cells were infected with this virus andsent for purification.

Purification of FL Human AKT1:

For the purification of full-length AKT1, 130 g sf9 cells (batch#41646W02) were resuspended in lysis buffer (buffer A, 1 L, pH 7.5)containing 25 mM HEPES, 100 mM NaCl, and 20 mM imidazole. The cell lysiswas carried out by Avestin (2 passes at 15K-20K psi). Cell debris wasremoved by centrifuging at 16K rpm for 1 hour and the supernatant wasbatch bound to 10 ml Nickel Sepharose HP beads at 4 C for over night.The beads were then transferred to column and the bound material waseluted with buffer B (25 mM HEPES, 100 mM NaCl, 300 mM imidazole, pH7.5). AKT eluting fractions were pooled and diluted 3 fold using bufferC (25 mM HEPES, 5 mM DTT; pH 7.5). The sample was filtered andchromatographed over a 10 mL Q-HP column pre-equilibrated with buffer Cat 2 mL/min.

The Q-HP column was washed with 3 column volume (CV) of buffer C, thenstep eluted with 5 CV 10% D, 5 CV 20% D, 5 CV 30% D, 5 CV 50% D and 5 CVof 100% D; where buffer D is 25 mM HEPES, 1000 mM NaCl, 5 mM DTT; pH7.5. 5 mL fractions collected. AKT containing fractions were pooled andconcentrated to 5 ml. The protein was next loaded to a 120 ml Superdex75 sizing column that was pre-equilibrated with 25 mM HEPES, 200 mMNaCl, 5 mM DTT; pH 7.5. 2.5 mL fractions were collected.

AKT 1 eluting fractions were pooled, aliquoted (1 ml) and stored at −80C. Mass spec and SDS-PAGE analysis were used to confirm purity andidentity of the purified full-length AKT1.

Full-length (FL) AKT2 and (FL) AKT3 were isolated and purified in asimilar fashion.

Full-Length AKT Enzyme Assay

Compounds of the present invention were tested for AKT 1, 2, and 3protein serine kinase inhibitory activity in substrate phosphorylationassays. This assay examines the ability of small molecule organiccompounds to inhibit the serine phosphorylation of a peptide substrate.The substrate phosphorylation assays use the catalytic domains of AKT 1,2, or 3. The method measures the ability of the isolated enzyme tocatalyze the transfer of the gamma-phosphate from ATP onto the serineresidue of a biotinylated synthetic peptide SEQ. ID NO: 1(Biotin-ahx-ARKRERAYSFGHHA-amide). Substrate phosphorylation wasdetected by the following procedure.

Assays were performed in 384well U-bottom white plates. 10 nM activatedAKT enzyme was incubated for 40 minutes at room temperature in an assayvolume of 20 ul containing 50 mM MOPS, pH 7.5, 20 mM MgCl2, 4 uM ATP, 8uM peptide, 0.04 uCi [g-33P] ATP/well, 1 mM CHAPS, 2 mM DTT, and 1 ul oftest compound in 100% DMSO. The reaction was stopped by the addition of50 ul SPA bead mix (Dulbecco's PBS without Mg²⁺ and Ca²⁺, 0.1% TritonX-100, 5 mM EDTA, 50 uM ATP, 2.5 mg/ml Streptavidin-coated SPA beads.)The plate was sealed, the beads were allowed to settle overnight, andthen the plate was counted in a Packard Topcount MicroplateScintillation Counter (Packard Instrument Co., Meriden, Conn.). The datafor dose responses were plotted as % Control calculated with the datareduction formula 100*(U1−C2)/(C1−C2) versus concentration of compoundwhere U is the unknown value, C1 is the average control value obtainedfor DMSO, and C2 is the average control value obtained for 0.1M EDTA.Data are fitted to the curve described by: y=((Vmax*x)/(K+x))

where Vmax is the upper asymptote and K is the IC50.

Compounds of the invention are tested for activity against AKT1, AKT2,and AKT3 in one or more of the above assays.

The compounds of the Examples were tested generally according to theabove AKT enzyme assays and in at least one experimental run exhibited apIC50 value: ≧5.5 against full length AKT1.

The compound of Example 2 was tested generally according to the aboveAKT enzyme assays and in at least one experimental run exhibited a pIC50value of 8.0 against full length AKT1.

In the above data, pIC50 is defined as −log(IC50) where the IC50 valueis expressed in molar units.

The pharmaceutically active compounds within the scope of this inventionare useful as AKT inhibitors in mammals, particularly humans, in needthereof.

The present invention therefore provides a method of treating cancer,arthritis and other conditions requiring AKT inhibition, which comprisesadministering an effective amount of a compound of Formula (I) orFormula (II) or a pharmaceutically acceptable salt, hydrate, solvate orpro-drug thereof. The compounds of Formula (I) and Formula (II) alsoprovide for a method of treating the above indicated disease statesbecause of their demonstrated ability to act as Akt inhibitors. The drugmay be administered to a patient in need thereof by any conventionalroute of administration, including, but not limited to, intravenous,intramuscular, oral, subcutaneous, intradermal, and parenteral.

The pharmaceutically active compounds of the present invention areincorporated into convenient dosage forms such as capsules, tablets, orinjectable preparations. Solid or liquid pharmaceutical carriers areemployed. Solid carriers include, starch, lactose, calcium sulfatedihydrate, terra alba, sucrose, talc, gelatin, agar, pectin, acacia,magnesium stearate, and stearic acid. Liquid carriers include syrup,peanut oil, olive oil, saline, and water. Similarly, the carrier mayinclude any prolonged release material, such as glyceryl monostearate orglyceryl distearate, alone or with a wax. The amount of solid carriervaries widely but, preferably, will be from about 25 mg to about 1 g perdosage unit. When a liquid carrier is used, the preparation will, forexample, be in the form of a syrup, elixir, emulsion, soft gelatincapsule, sterile injectable liquid such as an ampoule, or an aqueous ornonaqueous liquid suspension.

The pharmaceutical preparations are made following conventionaltechniques of a pharmaceutical chemist involving mixing, granulating,and compressing, when necessary, for tablet forms, or mixing, fillingand dissolving the ingredients, as appropriate, to give the desired oralor parenteral products.

Doses of the presently invented pharmaceutically active compounds in apharmaceutical dosage unit as described above will be an efficacious,nontoxic quantity preferably selected from the range of 0.001-100 mg/kgof active compound, preferably 0.001-50 mg/kg. When treating a humanpatient in need of an Akt inhibitor, the selected dose is administeredpreferably from 1-6 times daily, orally or parenterally. Preferred formsof parenteral administration include topically, rectally, transdermally,by injection and continuously by infusion. Oral and/or parenteral dosageunits for human administration preferably contain from 0.05 to 3500 mgof active compound.

Optimal dosages to be administered may be readily determined by thoseskilled in the art, and will vary with the particular Akt inhibitor inuse, the strength of the preparation, the mode of administration, andthe advancement of the disease condition. Additional factors dependingon the particular patient being treated will result in a need to adjustdosages, including patient age, weight, diet, and time ofadministration.

The method of this invention of inducing Akt inhibitory activity inmammals, including humans, comprises administering to a subject in needof such activity an effective Akt inhibiting amount of apharmaceutically active compound of the present invention.

The invention also provides for the use of a compound of Formula (I) orFormula (II) in the manufacture of a medicament for use as an Aktinhibitor.

The invention also provides for the use of a compound of Formula (I) orFormula (II) in the manufacture of a medicament for use in therapy.

The invention also provides for the use of a compound of Formula (I) orFormula (II) in the manufacture of a medicament for use in treatingcancer.

The invention also provides for the use of a compound of Formula (I) orFormula (II) in the manufacture of a medicament for use in treatingarthritis.

The invention also provides for a pharmaceutical composition for use asan Akt inhibitor which comprises a compound of Formula (I) or Formula(II) and a pharmaceutically acceptable carrier.

The invention also provides for a pharmaceutical composition for use inthe treatment of cancer which comprises a compound of Formula (I) orFormula (II) and a pharmaceutically acceptable carrier.

The invention also provides for a pharmaceutical composition for use intreating arthritis which comprises a compound of Formula (I) or Formula(II) and a pharmaceutically acceptable carrier.

No unacceptable toxicological effects are expected when compounds of theinvention are administered in accordance with the present invention.

In addition, the pharmaceutically active compounds of the presentinvention can be co-administered with further active ingredients, suchas other compounds known to treat cancer or arthritis, or compoundsknown to have utility when used in combination with an Akt inhibitor.

Without further elaboration, it is believed that one skilled in the artcan, using the preceding description, utilize the present invention toits fullest extent. The following Examples are, therefore, to beconstrued as merely illustrative and not a limitation of the scope ofthe present invention in any way.

Experimental Details

The compounds of Examples 1 to 47 are readily made according to Schemes1 to 3 or by analogous methods.

Preparation 1 Preparation of1,1-dimethylethyl(2-amino-2-phenylethyl)carbamate

a) 1,1-dimethylethyl(2-hydroxy-2-phenylethyl)carbamate

To a solution of 2-amino-1-phenylethanol (5 g, 36.4 mmol) in THF (182mL) at 25° C. was added Boc₂O (8.7 g, 40.1 mmol) in one portion. After0.5 h, the solution was concentrated and the residue used directlywithout further purification: LC-MS (ES) m/z=238 (M+H)⁺.

b)1,1-dimethylethyl[2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-2-phenylethyl]carbamate

To a solution of 1,1-dimethylethyl(2-hydroxy-2-phenylethyl)carbamate (2g, 8.44 mmol), phthalimide (1 g, 7.03 mmol) and triphenylphosphine (2.76g, 10.5 mmol) in THF (35 mL) at 25° C. was added DEAD (1.7 mL, 10.5mmol) dropwise. After 0.5 h, the solution was concentrated and purifiedvia column chromatography (silica, 15% EtOAc in hexanes) affording thetitle compound (2 g, 80%) as a white foam: LC-MS (ES) m/z=367 (M+H)⁺.

c) 1,1-dimethylethyl(2-amino-2-phenylethyl)carbamate

A solution of1,1-dimethylethyl[2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-2-phenylethyl]carbamate(2 g, 5.46 mmol) and either MeNH₂ (40 wt % in H₂O, 10 eq.) or NH₂NH₂ (10eq.) in MeOH (0.5M, 10 mL) was heated to 60° C. in a sealed tube. After12 h, the solution was concentrated and purified via columnchromatography (silica-dry load, 2% MeOH in DCM (1% NH₄OH)) affordingthe title compound (1.1 g, 85%) as a white solid: LC-MS (ES) m/z=237(M+H)⁺.

Preparation 2

Preparation of 1,1-dimethylethyl(2-amino-3-phenylpropyl)carbamate a)1-amino-3-phenyl-2-propanol

A solution of 2-(phenylmethyl)oxirane (7.5 g, 56.3 mmol) in NH₄OH (100mL) was stirred at 25° C. in a sealed tube. After 12 h, the solution wasconcentrated and used directly: LCMS (ES) m/e 152 (M+H)⁺.

b) 1,1-dimethylethyl(2-hydroxy-3-phenylpropyl)carbamate

To a solution of 1-amino-3-phenyl-2-propanol (7.6 g, 50 mmole) in THF(50 mL) at RT was added (Boc)₂O (12.0 g, 55 mmole). After stirring at RTfor 2 h, the reaction solution was concentrated under vacuum and theresidue purified on silica gel (5% MeOH in DCM (0.5% NH₄OH)) affordingthe title compound (13.1 g, 91%) as a clear yellow oil: LCMS (ES) m/z252 (M+H)⁺.

c)1,1-dimethylethyl[2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-3-phenylpropyl]carbamate

To a solution of 1,1-dimethylethyl(2-hydroxy-3-phenylpropyl)carbamate(10.0 g, 39.8 mmol), PPh₃ (12.5 g, 47.8 mmol) and phthalimide (6.44 g,43.8 mmol) in THF (125 mL) at RT was added DEAD (9.4 mL, 59.7 mmol) over5 min. After 1 h at RT, the reaction solution was concentrated andpurified on silica (hexanes/EtOAc, 2:1) to give the title compound as awhite solid (12.6 g, 83%): LCMS (ES) m/z 381 (M+H)⁺.

c) 1,1-dimethylethyl(2-amino-3-phenylpropyl)carbamate

NH₂NH₂ (12.5 mL, 394 mmol) was added to a THF/MeOH (50 mL/50 mL)solution of 1,1-dimethylethyl(2-amino-4-phenylbutyl)carbamate (7.5 g,19.7 mmol) and stirred at 50° C. in a sealed system. After 12 hours, thesolids were filtered, washing with methanol. The filtrate wasconcentrated and purified by column chromatography using 5% MeOH inCHCl₃containing 0.5% NH₄OH to give the title compound (3.75 g, 76%) as awhite solid: LC-MS (ES) m/z=251 (M+H)⁺.

Preparation 3

Preparation of2-{(2S)-2-amino-3-[2-(trifluoromethyl)phenyl]propyl}-1H-isoindole-1,3(2H)-dionea)1,1-dimethylethyl((1S)-2-hydroxy-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)carbamate

To a solution ofN-{[(1,1-dimethylethypoxy]carbonyl}-2-(trifluoromethyl)-L-phenylalanine(5 g, 15 mmol) in THF (75 mL) at 0° C. stirred was added BH₃-THF (45 mL,45 mmol-1M in THF). After 12 h, the reaction was quenched with AcOH:MeOH(1:5, 24 mL) and partitioned between saturated aqueous NaHCO₃ and DCM.The aqueous phase was then extracted several times with DCM. Thecombined organic fractions were dried over Na₂SO₄ and used directly (4.2g, 88%): LCMS (ES) m/e 320 (M+H)⁺.

b)1,1-dimethylethyl((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)carbamate

To a solution of1,1-dimethylethyl((1S)-2-hydroxy-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)carbamate(4.2 g, 13.2 mmol), triphenylphosphine (4.5 g, 17.1 mmol) andphthalimide (1.9 g, 13.2 mmol) in THF (66 mL) at 25° C. was addeddiethyl azodicarboxylate (2.7 mL, 17.1 mmol). After stirring at RT for 1h, the reaction solution was concentrated under vacuum and the residuepurified on silica gel (1% MeOH in DCM) affording the title compound(3.2 g, 54%) as a white solid: LCMS (ES) m/z 449 (M+H)⁺.

c)1,1-dimethylethyl{(2S)-2-amino-3-[2(trifluoromethyl)phenyl]propyl}carbamate

To a solution of1,1-dimethylethyl((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)carbamate(3.2 g, 7.1 mmol) in MeOH (35 mL) at RT was added 4M HCl in dioxane (18mL). After 12 h, the solution was concentrated affording the titlecompound (2.7 g, quant.) as the HCl salt: LCMS (ES) m/z 349 (M+H)⁺.

Preparation 4

Preparation of5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole

To a solution of 1-methyl pyrazole (4.1 g, 50 mmole) in THF (100 mL) at0° C. was added n-BuLi (2.2M in THF, 55 mmole). The reaction solutionwas stirred for 1 hour at RT and then cooled to −78° C. [J. HeterocyclicChem. 41, 931 (2004)]. To the reaction solution was added2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (12.3 mL, 60mmole). After 15 min at −78° C., the reaction was allowed to warm to 0°C. over 1 hour. The reaction was diluted with saturated NH₄Cl solutionand extracted with DCM. The organics were dried over Na₂SO₄ andconcentrated under vacuum to afford a tan solid (8.0 g, 77%) which wasused without further purification. LCMS (ES) m/z 127 (M+H)⁺ for[RB(OH)₂]; ¹H NMR (CDCl₃, 400 MHz) δ 7.57 (s, 1H), 6.75 (s, 1H), 4.16(s, 3H), and 1.41 (s, 12H).

Preparation 5

Preparation of3-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-(phenylmethyl)propanoicacid a) diethyl2-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl]-2-(phenylmethyl)butanedioate

To a suspension of NaH (2.2 g, 54 mmole) in THF (200 mL) was addeddiethyl 2-(phenylmethyl)butanedioate (12.5 g, 49.9 mmole). After 30 minat RT, bromomethyl phthalimide was added to the reaction mixture and thecontents were stirred for 14 h at RT. The reaction was quenched with H₂O(15 mL), diluted with Et₂O (300 mL) and layers separated. The organiclayer was concentrated under vacuum and the resulting residuerecrystallized from EtOH)(0°) to give the title compound (13 g, 64%) asa white solid: LCMS (ES) m/z 410 (M+H)⁺.

b) 3-amino-2-(phenylmethyl)propanoic acid

A solution of diethyl2-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl]-2-(phenylmethyl)butanedioate(13 g, 31.8 mmole) in conc. HCl (125 mL) and HOAc (30 mL) was sealed ina 1 L screwcap reaction vessel and the contents heated to 120° C. for 48h. The reaction solution was concentrated under vacuum and the resultingsolids washed with Et₂O affording the title compound (quant.) as a whitesolid: LCMS (ES) m/z 181(M+H)⁺.

c) 3-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-(phenylmethyl)propanoicacid

To a solution of 3-amino-2-(phenylmethyl)propanoic acid (from previousreaction) in dioxane (100 mL) and H₂O (20 mL) was added Boc anhydride(10.1 g, 46.5 mmole) and 6M NaOH solution (26 mL). After stirring at RTfor 12 h, the reaction solution was concentrated under vacuum,neutralized with 3M HCl and extracted with DCM. The organic layer wasdried over Na₂SO₄ and concentrated to a viscous oil. The oil was washedwith pentane and dried under high vacuum to give the title compound (5.0g, 17.9 mmole, 58% for 2 steps) as a white solid: LCMS (ES) m/z 280(M+H)⁺.

Preparation 6

Preparation of 5-iodo-1-methyl-1H-1,2,4-triazole

1-methyl-1H-1,2,4-triazole (2.05 g, 24.7 mmol) was added slowly over 15minutes to an Et₂O solution of nBuLi at −70° C. The mixture was stirredfor 60 minutes at −70° C. and allowed to warm to −30° C. A solution ofI₂ (6.5 g, 25.6 mmol) in THF (27 mL) was added slowly over 15 minutesand the mixture was allowed to warm to room temperature and stir for 60minutes. The mixture was partitioned with saturated Na₂S₂O₃, the phaseswere separated and the organic solvent removed. The crude iodide wasused without further purification: LCMS (ES) m/z 210 (M+H)⁺.

Example 1

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamidea)1,1-dimethylethyl(2-{[(5-bromo-2-pyridinyl)carbonyl]amino}-3-phenylpropyl)carbamate

To a solution of 5-bromo-2-pyridinecarboxylic acid (0.52 g, 2.6 mmol),PyBrOP (1.4 g, 3.0 mmol) and diisopropylethyl amine (1.0 mL, 6.0 mmol)in DCM (70 mL) at 25° C. was added1,1-dimethylethyl(2-amino-3-phenylpropyl)carbamate (0.5 g, 2.0 mmol).After 16 h, the solution was partitioned between H₂O and washed withDCM. The combined organic fraction were dried (Na₂SO₄), concentrated andpurified via column chromatography (silica, hexanes/EtOAc, 1:1)affording the title compound (0.60 g, 69%) as a white solid: LC-MS (ES)m/z=435 (M+H)⁺.

b)1,1-dimethylethyl[2-({[5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinyl]carbonyl}amino)-3-phenylpropyl]carbamate

To a solution of1,1-dimethylethyl(2-{[(5-bromo-2-pyridinyl)carbonyl]amino}-3-phenylpropyl)carbamate(0.58 g, 1.34 mmol) in dioxane/H₂O (5:1, 55 mL) was added K₂CO₃ (0.55 g,4.0 mmol), tetrakistriphenylphosphine Pd(0) (0.15 mg, 0.13 mmol), and5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole (0.39 g,2.0 mmol). The reaction mixture was heated to 80° C. in a sealed tubefor 12 h. The reaction solution was poured onto H₂O (100 mL) andextracted with DCM. The organics were dried (Na₂SO₄), concentrated undervacuum, and purified on silica gel (DCM/MeOH, 95:5) to give the titlecompound (0.39 g, 67%) as a white solid: LC-MS (ES) m/z=436 (M+H)⁺.

c)N-[2-amino-1-(phenylmethyl)ethyl]-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide

1,1-dimethylethyl[2-({[5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinyl]carbonyl}amino)-3-phenylpropyl]carbamate(0.39 g, 0.90 mmol) was dissolved in MeOH (10 mL) and THF (5 mL) andtreated with 4M HCl in dioxane (5 mL). After 3 h, the solution wasconcentrated affording the HCl salt of the title compound (0.32 g, 90%)as a white powder: LC-MS (ES) m/z=336 (M+H)⁺)⁺, ¹H NMR (d₄-MeOH, 400MHz) δ 8.87 (s, 1H), 8.22 (m, 2H), 7.81 (s, 1H), 7.32 (m, 4H), 7.24 (m,1H), 6.73 (s, 1H), 4.62 (m, 1H), 4.00 (s, 1H), 3.27 (m, 2H), 3.05 (d,J=7.3 Hz, 2H).

Example 2

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-6-(1-methyl-1H-pyrazol-5-yl)-3-pyridinecarboxamide

The title compound was prepared as a white solid according to theexample of Example 1, except substituting 2-bromo-5-pyridinecarboxylicacid (0.52 g, 2.6 mmol) for 5-bromo-2-pyridinecarboxylic acid: LC-MS(ES) m/z 336 (M+H)⁺, ¹H NMR (d₄-MeOH, 400 MHz) δ 9.10 (s, 1H), 8.43 (d,J=8.3 Hz, 1H), 8.04 (d, J=8.3 Hz, 1H), 7.82 (d, J=2.3 Hz, 1H), 7.35 (m,4H), 7.30 (m, 1H), 7.01 (s, 1H), 4.59 (m, 1H), 4.39 (s, 3H), 3.26 (m,2H), 3.05 (m, 2H).

Example 3

Preparation ofN-(2-amino-1-benzylethyl)-5-(1-methyl-1H-pyrazol-5-yl)pyrimidine-2-carboxamidea) 5-(1-methyl-1H-pyrazol-5-yl)pyrimidine-2-carboxylic acid

To a solution of1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(1.35 g, 6.5 mmol) in dioxane/H₂O (5:1, 59 mL) was added K₂CO₃ (2.8 g,20.3 mmol), tetrakistriphenylphosphine Pd(0) (340 mg, 0.3 mmol), and5-bromo-2-pyrimidinecarboxylic acid (1.2 g, 5.9 mmol). The reactionmixture was heated to 80° C. in a sealed tube for 12 h. The reactionsolution was poured onto H₂O (100 mL) and extracted with DCM. Theorganics were dried (Na₂SO₄), concentrated under vacuum to give thetitle compound (0.9 g, 36%) as a tan solid: LC-MS (ES) m/z=437 (M+H)⁺.

b)1,1-dimethylethyl[2-({[5-(1-methyl-1H-pyrazol-5-yl)-2-pyrimidinyl]carbonyl}amino)-3-phenylpropyl]carbamate

To a solution of 5-(1-methyl-1H-pyrazol-5-yl)pyrimidine-2-carboxylicacid (194 mg, 0.4 mmol), PyBrOP (330 mg, 0.71 mmol) and diisopropylethylamine (0.27 mL, 1.5 mmol) in DCM (7 mL) at 25° C. was added1,1-dimethylethyl(2-amino-3-phenylpropyl)carbamate (174 mg, 0.7 mmol).After 16 h, the solution was concentrated and purified via columnchromatography (silica, hexanes/EtOAc, 2:1) affording the title compound(76 mg, 39%) as a white solid: LC-MS (ES) m/z=437 (M+H)⁺.

c)N-(2-amino-1-benzylethyl)-5-(1-methyl-1H-pyrazol-5-yl)pyrimidine-2-carboxamide

1,1-dimethylethyl[2-({[5-(1-methyl-1H-pyrazol-5-yl)-2-pyrimidinyl]carbonyl}amino)-3-phenylpropyl]carbamate(76 mg, 0.17 mmol) was dissolved in MeOH (1 mL) and CHCl₃ (10 mL) andtreated with 4M HCl in dioxane (5 mL). After 3 h, the solution wasconcentrated and purified via column chromatography (silica, CHCl₃/MeOH,90:10 containing 1% NH₄OH affording the free base of the title compound(40 mg, 68%) as a white powder.

The HCl salt was made by addition of an excess of 1M HCl in Et₂O toyield the final compound as the HCl salt: LC-MS (ES) m/z=337 (M+H)⁺, ¹HNMR (d₄-MeOH, 400 MHz) δ ppm 2.91-3.01 (m, 4H) 4.00 (s, 3H) 4.37-4.46(m, 1H) 6.69 (d, J=2.02 Hz, 1H) 7.18 (t, J=7.20 Hz, 1H) 7.23-7.33 (m,4H) 7.62 (d, J=2.02 Hz, 1 H) 9.11 (s, 2H).

Example 4

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to theprocedure of Example 3, except substituting 4-bromobenzoic acid (102 mg,0.5 mmol) for 5-bromo-2-pyrimidinecarboxylic acid: LC-MS (ES) m/z=335(M+H)⁺, ¹H NMR (d₄-MeOH, 400 MHz) δ 7.96 (d, J=12 Hz, 2H), 7.63 (d, J=12Hz, 2H), 7.4 (m, 1H), 7.34 (m, 4H), 7.23 (m, 1H), 6.58 (s, 1H), 4.63 (m,1H), 3.96 (s, 3H), 3.25 (m, 2H), 3.05 (m, 2H).

Example 5

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-2-chloro-4-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to theprocedure of Example 3, except substituting 4-bromo-2-chlorobenzoic acid(118 mg, 0.5 mmol) for 5-bromo-2-pyrimidinecarboxylic acid: LC-MS (ES)m/z=369 (M+H)⁺, ¹H NMR (d₄-MeOH, 400 MHz) δ 7.70 (s, 1H), 7.65 (m, 1H),7.54 (m, 2H), 7.36 (m, 4H), 7.29 (m, 1H), 6.57 (m, 1H), 4.63 (m, 1H),4.89 (s, 3H), 3.21 (m, 2H), 3.08 (m, 1H), 2.98 (m, 1H).

Example 6

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-3-chloro-4-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a yellow solid according to theprocedure of Example 3, except substituting 4-bromo-3-chlorobenzoic acid(118 mg, 0.5 mmol) for 5-bromo-2-pyrimidinecarboxylic acid: LC-MS (ES)m/z=369 (M+H)⁺)⁺, ¹H NMR (d₄-MeOH, 400 MHz) δ 8.03 (s, 1H), 7.9 (m, 2H),7.6 (m, 1H), 7.38 (m, 1H), 7.32 (m, 1H), 7.25 (m, 1H), 6.64 (m, 1H),4.62 (m, 1H), 3.83 (s, 3H), 3.24 (m, 2H), 3.04 (m, 2H).

Example 7

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-pyrazol-5-yl)-3-(trifluoromethyl)benzamidea)1,1-dimethylethyl[2-({[4-hydroxy-3-(trifluoromethyl)phenyl]carbonyl}amino)-3-phenylpropyl]carbamate

To a solution of 4-hydroxy-3-trifluoromethylbenzoic acid (0.54 mg, 2.6mmol), PyBrOP (0.77 g, 6.0 mmol) and diisopropylethyl amine (1 mL, 6.0mmol) in DCM (50 mL) at 25° C. was added1,1-dimethylethyl(2-amino-3-phenylpropyl)carbamate (0.50 mg, 2.0 mmol).After 12 h, the solution was concentrated and purified on silica(EtOAc/hexanes, 2:1) to give the title compound (0.19 g, 17%) as acolorless oil: LC-MS (ES) m/z=439 (M+H)⁺.

b)4-({[2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-1-(phenylmethyl)ethyl]amino}carbonyl)-2-(trifluoromethyl)phenyltrifluoromethanesulfonate

To a solution of1,1-dimethylethyl[2-({[4-hydroxy-3-(trifluoromethyl)phenyl]carbonyl}amino)-3-phenylpropyl]carbamate(0.18 g, 0.41 mmoles) in DCM (15 mL) at room temperature was addedtriethylamine (0.17 mL, 1.2 mmol) and N-phenyltrifluoromethanesulfonimide (0.19 g, 0.53 mmoles). After 24 h, the solution wasconcentrated and purified via column chromatography (silica,hexanes/EtOAc, 4:1) yielding the the tilte compound (100 mg, 43%) as awhite solid: LC-MS (ES) m/z=571 (M+H)⁺.

c)1,1-dimethylethyl[2-({[4-(1-methyl-1H-pyrazol-5-yl)-3-(trifluoromethyl)phenyl]carbonyl}amino)-3-phenylpropyl]carbamate

To a solution of4-({[2-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-1-(phenylmethyl)ethyl]amino}carbonyl)-2-(trifluoromethyl)phenyltrifluoromethanesulfonate (0.21 g, 0.37 mmol) in dioxane/H₂O (5:1, 10mL) was added K₂CO₃ (150 mg, 1.1 mmol), tetrakistriphenylphosphine Pd(0)(46 mg, 37 umol) and5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole (140 mg,0.74 mmol). The reaction mixture was heated to 80° C. in a sealed tubefor 12 h. The reaction solution was poured onto H₂O (100 mL) andextracted with DCM. The organics were dried (Na₂SO₄), concentrated undervacuum and purified on silica (EtOAc) to give the title compound (0.16g, 86%) as a white solid: LC-MS (ES) m/z=503 (M+H)⁺.

d)N-[2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-pyrazol-5-yl)-3-(trifluoromethyl)benzamide

1,1-dimethylethyl[2-({[4-(1-methyl-1H-pyrazol-5-yl)-3-(trifluoromethyl)phenyl]carbonyl}amino)-3-phenylpropyl]carbamate(0.16 g, 0.32 mmoles) was dissolved in MeOH (10 mL) and THF (3 mL) andtreated with excess 4M HCl in dioxane. After 2 h at RT, the reactionsolution was concentrated under vacuum affording the HCl salt of thetitle compound (150 mg, quant.) as a white solid: LC-MS (ES) m/z=403(M+H)⁺, ¹H NMR (MeOD, 400 MHz) δ 8.29 (s, 1H), 8.17 (d, J=7.8 Hz, 1H),7.84 (s, 1H), 7.34 (m, 4H), 7.25 (m, 1H), 6.53 (s, 1H), 4.65 (m, 1H),3.73 (s, 3H), 3.26 (m, 2H) and 3.04 (m, 2H).

Example 8

Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamidea)5-bromo-N-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-2-pyridinecarboxamide

To a solution of 5-bromo-2-pyridinecarboxylic acid (97 mg, 2.6 mmol),PyBrOP (269 mg, 0.578 mmol) and diisopropylethyl amine (419 uL, 2.41mmol) in DCM (5 mL) at 25° C. was added2-{(2S)-2-amino-3-[2-(trifluoromethyl)phenyl]propyl}-1H-isoindole-1,3(2H)-dione(168 mg, 0.481 mmol)[prepared in preparation 3]. After 16 h, thesolution was partitioned between H₂O and washed with DCM. The combinedorganic fraction were dried (Na₂SO₄), concentrated and used directlywithout further purification: LC-MS (ES) m/z=435 (M+H)⁺.

b)1,1-dimethylethyl{(2S)-2-{[(5-bromo-2-pyridinyl)carbonyl]amino}-3-[2-(trifluoromethyl)phenyl]propyl}carbamate

i) To a solution of5-bromo-N-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-2-pyridinecarboxamide(crude from part a) in MeOH-THF (1:1, 5 mL) was added hydrazine (151 uL,4.80 mmol) at 25° C. After 12 h, the solution was filtered and thefiltrate was concentrated. The resulting residue was purified via columnchromatography (silica, 2% MeOH in DCM (1% NH₄OH)) affording the freebase (192 mg, quant.) of the title compound: LCMS (ES) m/e 402 (M+H)⁺.

ii) The free base (192 mg, 0.478 mmol) was dissolved in THF (6 mL) andtreated with Boc₂O (144 mg, 0.669 mmol). After 30 min, the solution wasconcentrated affording the title compound (300 mg, quant.) as a clearoil which was used directly: LCMS (ES) m/e 502 (M+H)⁺.

c)1,1-dimethylethyl{(2S)-2-({[5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinyl]carbonyl}amino)-3-[2-(trifluoromethyl)phenyl]propyl}carbamate

To a solution of1,1-dimethylethyl{(2S)-2-{[(5-bromo-2-pyridinyl)carbonyl]amino}-3-[2-(trifluoromethyl)phenyl]propyl}carbamate(300 mg, 0.60 mmol) in dioxane/H₂O (5:1, 6 mL) was added K₂CO₃ (331 mg,2.4 mmol), tetrakistriphenylphosphine Pd(0) (35 mg, 29.9 mmol), and5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole (162 mg,0.78 mmol). The reaction mixture was heated to 80° C. in a sealed tubefor 12 h. The reaction solution was poured onto H₂O (100 mL) andextracted with DCM. The organics were dried (Na₂SO₄) and concentratedunder vacuum affording the title compound which was used directlywithout further purification: LC-MS (ES) m/z=504 (M+H)⁺.

d)N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide

1,1-dimethylethyl{(2S)-2-({[5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinyl]carbonyl}amino)-3-[2-(trifluoromethyl)phenyl]propyl}carbamate(crude from part c) was dissolved in DCM (2 mL) and treated with TFA (1mL). After 30 min., the solution was concentrated and neutralizedthrough a plug of silica (2% MeOH in DCM (1% NH4OH)) affording the freebase of the title compound.

The free base was dissolved in MeOH (2 mL) and treated with excess 4MHCl in dioxane (500 uL) affording the HCl salt of the title compound(110 mg, 45%-3 steps) as a white solid: LCMS (ES) m/z=404 (M+H)⁺, ¹H NMR(400 MHz, DMSO-d₆) δ ppm 9.07 (d, J=9.60 Hz, 1H) 8.85 (d, J=1.52 Hz, 1H)8.18 (dd, J=8.08, 2.27 Hz, 1H) 8.03 (d, J=8.08 Hz, 4H) 7.69 (d, J=7.83Hz, 1H) 7.57 (d, J=2.02 Hz, 1H) 7.47-7.54 (m, 2H) 7.41 (dd, J=7.96, 2.15Hz, 1H) 6.65 (d, J=2.02 Hz, 1H) 4.51-4.55 (m, 1H) 3.92 (s, 3H) 3.13-3.16(m, 4H).

Example 9

Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-chloro-4-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to theprocedure of Example 8, except substituting 4-bromo-3-chlorobenzoic acid(88 mg, 0.372 mmol) for 5-bromo-2-pyridinecarboxylic acid: LCMS (ES)m/z=437 (M+H)⁺, ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.88 (br. s., 1H) 8.08(bs, 4H) 7.87 (br. s., 1 H) 7.71 (d, J=7.83 Hz, 1H) 7.54-7.61 (m, 4H)7.44 (s, 1H) 6.40 (d, J=1.77 Hz, 1 H) 4.59 (d, J=4.80 Hz, 1H) 3.66 (s,3H) 3.06-3.17 (m, 4H).

Example 10

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-4-chloro-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamidea) 5-bromo-2-methylpyridine N-oxide

To a solution of 5-bromo-2-methylpyridine (1 g, 5.81 mmol) and ureahydrogen peroxide (1.1 g, 12.2 mmol) in DCM (30 mL) at 0° C. was addedtrifluoroacetic anhydride (1.6 mL, 11.6 mmol). After warming to 25° C.over 12 h, the solution was partitioned between H₂O-DCM. The aqueousphase was washed several times with DCM and the combined organicfractions were dried over Na₂SO₄, concentrated and purified via columnchromatography (silica, 1% MeOH in DCM (1% NH₄OH)) yielding the titlecompound (660 mg, 60%) as a white solid: LCMS (ES) m/z=190 (M+H)⁺.

b) 5-bromo-2-methyl-4-nitropyridine N-oxide

To a solution of 5-bromo-2-methylpyridine N-oxide (660 mg, 3.51 mmol) insulfuric acid (1.6 mL) at 0° C. was added a premixed solution of nitricacid (1.5 mL) in sulfuric acid (2.5 mL). The resulting reaction waswarmed to 25° C. then heated to 80° C. After 12 h, the solution waspoured into ice then, neutralized with 6N NaOH and extracted severaltimes with DCM. The combined organic fractions were dried over Na₂SO₄,concentrated and purified via column chromatography (silica, 1% MeOH inDCM) yielding the title compound (558 mg, 81%) as a yellow solid: LCMS(ES) m/z=235 (M+H)⁺.

c) 5-bromo-4-chloro-2-methylpyridine N-oxide

To a solution of 5-bromo-2-methyl-4-nitropyridine N-oxide (558 mg, 2.41mmol) in DCM (3 mL) at 0° C. was added dropwise a solution of POCl₃ (1.3mL, 7.22 mmol) in DCM (3.2 mL). After 12 h at 25° C. the solution wasadded to ice, adjusted to pH ˜8 and extracted several times with DCM.The combined organic fractions were dried over Na₂SO₄, concentrated andpurified via column chromatography (silica, 1.5% MeOH in DCM) yieldingthe title compound (426 mg, 80%) as a white solid: LCMS (ES) m/z=224(M+H)⁺.

d) 5-bromo-4-chloro-2-pyridinyl)methanol

To a solution of 5-bromo-2-methyl-4-nitropyridine N-oxide (426 mg, 1.93mmol) in DCM (5 mL) at 0° C. was added trifluoroacetic anhydride (817uL, 5.78 mmol). After warming to 25° C. over 12 h, the solution waspartitioned between 6N NaOH-DCM. The aqueous phase was washed severaltimes with DCM and the combined organic fractions were dried overNa₂SO₄, concentrated and purified via column chromatography (silica, 1%MeOH in DCM) yielding the title compound (187 mg, 44%) as a white solid:LCMS (ES) m/z=223 (M+H)⁺.

e) 5-bromo-4-chloro-2-pyridinecarboxylic acid

KMnO₄ (174 mg, 1.1 mmol) in H₂O was added dropwise to a solution of(5-bromo-4-chloro-2-pyridinyl)methanol (187 mg, 0.846 mmol) in 10%Na₂CO₃ (1 mL). After 2 h, the solution was filtered through Celite®,adjusted to pH ˜4 and the resulting precipitate was filtered thenazeotropically dried with toluene affording the title compound (135 mg,68%) as a white powder: LCMS (ES) m/z=237 (M+H)⁺.

f)N-[2-amino-1-(phenylmethyl)ethyl]-4-chloro-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide

The title compound was prepared as a white solid according to theexample of Example 1, except substituting5-bromo-4-chloro-2-pyridinecarboxylic acid (135 mg, 0.572 mmol) for5-bromo-2-pyridinecarboxylic acid: LCMS (ES) m/z=370 (M+H)⁺, ¹H NMR (400MHz, DMSO-d₆) δ ppm 9.07 (d, J=9.35 Hz, 1H) 8.75 (s, 1H) 8.14 (s, 1H)8.03 (br. s., 3H) 7.61 (d, J=2.02 Hz, 1H) 7.26 (d, J=3.28 Hz, 3H)7.23-7.30 (m, 1H) 7.17-7.22 (m, 1H) 6.53 (d, J=2.02 Hz, 1H) 4.53 (d,J=4.29 Hz, 1H) 3.71 (s, 3H) 2.96-3.17 (m, 4H).

Example 11

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide

The title compound was prepared as a white solid according to theprocedure of Example 1, except substituting 4-bromo-2-pyridinecarboxylicacid (300 mg, 1.48 mmol) for 5-bromo-2-pyridinecarboxylic acid: LCMS(ES) m/z=436 (M+H)⁺, ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.99 (d, J=9.09 Hz,1H) 8.77 (d, J=5.05 Hz, 1H) 8.08 (d, J=1.01 Hz, 1H) 8.01 (s, 3H) 7.84(dd, J=5.05, 1.77 Hz, 1 H) 7.57 (d, J=2.02 Hz, 1H) 7.23-7.30 (m, 4H)7.19 (td, J=6.19, 2.53 Hz, 1H) 6.71 (d, J=1.77 Hz, 1H) 4.52 (m, 1H) 3.96(s, 3H) 2.97-3.10 (m, 4H).

Example 12

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-6-chloro-5-(1-methyl-1H-pyrazol-5-yl-2-pyridinecarboxamidea) 3-bromo-2-chloro-6-methylpyridine N-oxide

To a solution of 3-bromo-2-chloro-6-methylpyridine (1 g, 4.84 mmol) andurea hydrogen peroxide (911 mg, 9.69 mmol) in DCM (24 mL) at 0° C. wasadded trifluoroacetic anhydride (1.4 mL, 9.69 mmol). After warming to25° C. over 12 h, the solution was partitioned between H₂O-DCM. Theaqueous phase was washed several times with DCM and the combined organicfractions were dried over Na₂SO₄, concentrated and purified via columnchromatography (silica, 1% MeOH in DCM) yielding the title compound (1.1g, quant.) as a white solid: LCMS (ES) m/z=223 (M+H)⁺.

b)N-[2-amino-1-(phenylmethyl)ethyl]-6-chloro-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide

The title compound was prepared as a white solid according to Example10, except substituting 3-bromo-2-chloro-6-methylpyridine N-oxide (1.1g, 4.95 mmol) for 5-bromo-4-chloro-2-methylpyridine N-oxide: LCMS (ES)m/z=370 (M+H)⁺, ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.83 (d, J=9.35 Hz, 1H)8.10-8.17 (m, 1H) 8.06 (d, J=7.83 Hz, 4H) 7.58 (d, J=2.02 Hz, 1H)7.24-7.32 (m, 4H) 7.18-7.23 (m, 1 H) 6.49 (d, J=1.77 Hz, 1H) 4.53 (d,J=3.79 Hz, 1H) 3.70 (s, 3H) 3.17 (s, 1H) 2.98 (d, J=7.83 Hz, 3H).

Example 13

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-6-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide

The title compound was prepared as a white solid according to theprocedure of Example 3, except substituting 6-bromo-2-pyridinecarboxylicacid (1.2 g, 5.9 mmol) for 5-bromo-2-pyrimidinecarboxylic acid: LC-MS(ES) m/z 336 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 2.98-3.09 (m, 4H) 4.07(s, 3H) 4.51 (ddd, J=7.89, 4.99, 2.53 Hz, 1H) 6.77 (d, J=2.02 Hz, 1H)7.17-7.22 (m, 1H) 7.23-7.30 (m, 4H) 7.54 (d, J=2.27 Hz, 1H) 7.89 (q,J=4.55 Hz, 1H) 8.04 (d, J=4.04 Hz, 2H).

Example 14

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-5-(1-methyl-1H-pyrazol-5-yl)-3-pyridinecarboxamide

The title compound was prepared as a white solid according to theprocedure of Example 3, except substituting 5-bromo-3-pyridinecarboxylicacid (2.1 g, 10.4 mmol) for 5-bromo-2-pyrimidinecarboxylic acid: LC-MS(ES) m/z 336 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 2.77-3.04 (m, 4H) 3.91(s, 3H) 4.39 (s, 1H) 6.54 (d, J=2.02 Hz, 1H) 7.16-7.23 (m, 1H) 7.25-7.31(m, 4H) 7.58 (d, J=2.02 Hz, 1H) 8.23 (t, J=2.15 Hz, 1H) 8.81 (d, J=2.27Hz, 1H) 8.92 (d, J=2.02 Hz, 1H).

Example 15

PreparationN-[2-amino-1-(phenylmethyl)ethyl]-2-(1-methyl-1H-pyrazol-5-yl)-4-pyridinecarboxamide

The title compound was prepared as a white solid according to theprocedure of Example 3, except substituting 2-bromo-4-pyridinecarboxylicacid (1.0 g, 5.2 mmol) for 5-bromo-2-pyrimidinecarboxylic acid: LC-MS(ES) m/z 336 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 2.84-2.94 (m, 2H)2.96-3.02 (m, 2H) 4.16 (s, 3H) 4.40 (ddd, J=8.34, 4.80, 2.27 Hz, 1H)6.75 (d, J=2.02 Hz, 1H) 7.19 (td, J=6.00, 2.65 Hz, 1H) 7.25-7.30 (m, 4H)7.53 (d, J=2.02 Hz, 1H) 7.60 (dd, J=5.18, 1.64 Hz, 1H) 7.95 (s, 1H) 8.73(d, J=5.31 Hz, 1H).

Example 16

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-3-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to theprocedure of Example 3, except substituting 3-bromo-benzoic acid (101mg, 0.5 mmol) for 5-bromo-2-pyrimidinecarboxylic acid: LC-MS (ES) m/z335 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 3.01-3.08 (m, 2H) 3.21-3.28 (m,2H) 4.00 (d, J=1.77 Hz, 3H) 4.60-4.67 (m, 1H) 6.71 (d, J=2.53 Hz, 1H)7.20-7.25 (m, 1H) 7.29-7.35 (m, 4H) 7.66 (t, J=7.71 Hz, 1H) 7.76 (dd,J=7.83, 1.26 Hz, 1H) 7.92-7.97 (m, 3H).

Example 17

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-3-methyl-4-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to theprocedure of Example 3, except substituting 4-bromo-3-methylbenzoic acid(108 mg, 0.5 mmol) for 5-bromo-2-pyrimidinecarboxylic acid: LC-MS (ES)m/z 349 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 2.28 (s, 3H) 3.02-3.08 (m,2H) 3.21-3.28 (m, 2H) 3.80 (s, 3H) 4.59-4.67 (m, 1H) 6.62-6.63 (m, 1H)7.22-7.27 (m, 1H) 7.30-7.36 (m, 4H) 7.43 (d, J=8.08 Hz, 1H) 7.76 (dd,J=7.83, 1.26 Hz, 1H) 7.82 (s, 1H) 8.01-8.03 (m, 1H).

Example 18

Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzamidea). 3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzoic acid

To a solution of 4-bromo-3-fluorobenzoic acid (110 mg, 0.5 mmol) indioxane/H₂O (5:1, 6 mL) was added K₂CO₃ (207 mg, 1.5 mmol),tetrakistriphenylphosphine Pd(0) (29 mg, 0.025 mmol) and5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole (107 mg,0.55 mmol). The reaction mixture was heated to 80° C. in a sealed tubefor 12 h and was then partitioned between 6N NaOH and DCM. The pH of theaqueous phase was adjusted to ˜3 with 3M HCl and washed several timeswith DCM. The combined organic fractions were dried (Na₂SO₄),concentrated under vacuum and used directly without further purification(98 mg, 89%): LC-MS (ES) m/z=221 (M+H)+.

b)N-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzamide

To a solution of 3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzoic acid (66mg, 0.3 mmol), PyBrOP (169 mg, 0.37 mmol) and diisopropylethyl amine(263 μL, 1.5 mmol) in DCM (3 mL) at 25° C. was added2-{(2S)-2-amino-3-[2-(trifluoromethyl)phenyl]propyl}-1H-isoindole-1,3(2H)-dione-HCl(105 mg, 0.3 mmol)[prepared according to Preparation 3]. After 16 h, thesolution was partitioned between H₂O and washed with DCM. The combinedorganic fractions were dried (Na₂SO₄), concentrated and purified viacolumn chromatography (silica) affording the title compound (112 mg,68%) as a white solid: LC-MS (ES) m/z=551 (M+H)+.

c)N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzamide

To a solution ofN-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzamide(112 mg, 0.2 mmol) in MeOH/THF (2 mL, 1:1) at RT was added hydrazine(0.12 mL, 3.93 mmol). After stirring for 18 h at RT, the reactionsolution was concentrated under vacuum and purified via columnchromatography (silica, 3% MeOH in DCM (1% NH4OH)) yielding the titlecompound.

The neutral compound from above was dissolved in MeOH (2 mL), treatedwith excess 4M HCl in dioxane (0.5 mL) and concentrated affording theHCl salt of the title compound as a white solid: LC-MS (ES) m/z 421(M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 3.12-3.20 (m, 1H) 3.24-3.32 (m, 3H)3.86 (d, J=1.26 Hz, 3 H) 4.72-4.79 (m, 1H) 6.55 (d, J=2.02 Hz, 1H)7.41-7.48 (m, 1H) 7.52-7.58 (m, 2H) 7.58-7.63 (m, 1H) 7.70-7.74 (m, 2H)7.74-7.82 (m, 2H).

Example 19

Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-methyl-4-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to theprocedure of Example 18, except substituting 4-bromo-3-methylbenzoicacid (109 mg, 0.5 mmol) for 4-bromo-3-fluorobenzoic acid: LC-MS (ES) m/z417 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 2.24 (s, 3H) 3.09-3.17 (m, 1H)3.21-3.29 (m, 3 H) 3.65 (s, 3H) 4.73 (s, 1H) 6.31-6.32 (m, 1H) 7.37 (d,J=8 Hz, 1H) 7.44-7.47 (m, 1H) 7.52-7.59 (m, 3H) 7.69-7.75 (m, 2H)7.78-7.80 (m, 1H).

Example 20

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-3-hydroxy-4-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a yellow solid according to theprocedure of Example 3, except substituting 4-bromo-3-hydroxybenzoicacid (108 mg, 0.5 mmol) for 5-bromo-2-pyrimidinecarboxylic acid: LC-MS(ES) m/z 351(M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 3.01 (s, 2H) 3.18 (s,2H) 3.76 (s, 3 H) 4.60 (s, 1H) 6.35 (s, 2H) 7.25 (s, 1H) 7.30 (s, 2H)7.32 (s, 4H) 7.37 (s, 1H) 7.57 (s, 1H).

Example 21

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to Example 3,except substituting 4-bromo-3-fluorobenzoic acid (110 mg, 0.5 mmol) for5-bromo-2-pyrimidinecarboxylic acid: LC-MS (ES) m/z 421 (M+H)⁺, ¹H NMR(400 MHz, MeOD) δ ppm 3.06 (d, J=7.07 Hz, 2H) 3.23-3.30 (m, 2H) 4.02 (s,3H) 4.60-4.68 (m, 1H) 6.89-6.91 (m, 1H) 7.24 (d, J=5.81 Hz, 1H)7.28-7.36 (m, 4H) 7.71 (t, J=7.33 Hz, 1H) 7.77-7.87 (m, 2H) 8.22-8.32(m, 1H).

Example 22

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-3-chloro-5-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to Example 3,except substituting 3-bromo-5-chlorobenzoic acid (282 mg, 1.2 mmol) for5-bromo-2-pyrimidinecarboxylic acid: LC-MS (ES) m/z 369 (M+H)⁺, ¹H NMR(400 MHz, MeOD) δ ppm 3.05 (d, J=7.83 Hz, 2H) 3.26 (d, J=6.57 Hz, 2H)4.09 (s, 3H) 4.59-4.67 (m, J=7.14, 7.14, 7.14, 7.14 Hz, 1H) 6.92 (d,J=2.78 Hz, 1H) 7.23 (t, J=6.82 Hz, 1H) 7.28-7.36 (m, 4H) 7.86 (s, 1H)8.00 (s, 2H) 8.20 (s, 1H).

Example 23

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-2-chloro-5-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to theprocedure of Example 3, except substituting 5-bromo-2-chlorobenzoic acid(282 mg, 1.2 mmol) for 5-bromo-2-pyridinecarboxylic acid: LC-MS (ES) m/z369 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 2.93-3.02 (m, 1H) 3.04-3.11 (m,1H) 3.15-3.27 (m, 2H) 4.04 (s, 3H) 4.60-4.68 (m, 1H) 6.82 (s, 1H)7.23-7.30 (m, 1H) 7.30-7.37 (m, 4H) 7.58-7.69 (m, 3H), 8.12 (s, 1H).

Example 24

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-4-chloro-3-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as an off-white solid according toExample 3, except substituting 3-bromo-4-chlorobenzoic acid (282 mg, 1.2mmol) for 5-bromo-2-pyridinecarboxylic acid: LC-MS (ES) m/z 369 (M+H)⁺,¹H NMR (400 MHz, MeOD) δ ppm 2.98-3.06 (m, 2H) 3.17-3.26 (m, 2H)3.79-3.83 (m, 3H) 4.59 (s, 1H) 6.59 (d, J=2.27 Hz, 1H) 7.23 (dt, J=6.00,2.94 Hz, 1H) 7.28-7.34 (m, 4H) 7.73 (d, J=8.59 Hz, 1H) 7.85-7.95 (m,3H).

Example 25

Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-chloro-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide

The title compound was prepared as a white solid according to Example 8,except substituting 5-bromo-4-chloro-2-pyridinecarboxylic acid (142 mg,0.6 mmol)[prepared according to Example 10] for 4-bromo-3-fluorobenzoicacid: LC-MS (ES) m/z 438 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 3.12-3.21(m, 1H) 3.30 (d, J=4.04 Hz, 3H) 3.80 (s, 3H) 4.73 (d, J=4.55 Hz, 1H)6.56-6.62 (m, 1H) 7.41 (t, J=7.07 Hz, 1H) 7.50 (t, J=7.33 Hz, 1H)7.53-7.59 (m, 1H) 7.70 (d, J=7.83 Hz, 1H) 7.73-7.79 (m, 1H) 8.20 (s, 1H)8.72 (s, 1H)

Example 26

Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-chloro-3-(1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to Example18, except substituting 3-bromo-4-chlorobenzoic acid (282 g, 1.2 mmol)for 4-bromo-3-fluorobenzoic acid: LC-MS (ES) m/z 437 (M+H)⁺, ¹H NMR (400MHz, MeOD) δ ppm 3.18 (d, J=9.85 Hz, 1H) 3.22-3.31 (m, 3H) 3.91 (d,J=7.33 Hz, 3H) 4.70-4.76 (m, 1H) 6.65-6.78 (m, 1H) 7.43 (t, J=7.45 Hz,1H) 7.50-7.58 (m, 2H) 7.71 (d, J=7.83 Hz, 1H) 7.77 (d, J=8.84 Hz, 1H)7.98-8.08 (m, 3H).

Example 27

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-3-fluorobenzamidea) 3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzoic acid

To a solution of 4-bromo-3-fluorobenzoic acid (220 mg, 1.0 mmol) indioxane/H₂O (5:1, 12 mL) was added K₂CO₃ (414 mg, 3.0 mmol),tetrakistriphenylphosphine Pd(0) (58 mg, 0.05 mmol) and5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole (214 mg,1.1 mmol). The reaction mixture was heated to 80° C. in a sealed tubefor 12 h and was then partitioned between 6N NaOH and DCM. The pH of theaqueous phase was adjusted to ˜3 with 3M HCl and washed several timeswith DCM. The combined organic fractions were dried (Na₂SO₄),concentrated under vacuum and used directly without further purification(196 mg, 89%): LC-MS (ES) m/z=221 (M+H)⁺.

b) 4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-3-fluorobenzoic acid

A solution of 3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzoic acid (196 mg,0.89 mmol) and N-chlorosuccinimide (119 mg, 0.89 mmol) in THF (5 mL) wasstirred in a sealed tube at 70° C. After 1 h, the solution waspartitioned between H₂O-DCM, the aqueous phase was adjusted to pH 3 andthe aqueous phase was washed several times with DCM. The combinedorganic fractions were dried (Na₂SO₄), concentrated under vacuum andused directly without further purification (192 mg, 85%): LC-MS (ES)m/z=255 (M+H)⁺.

c).1,1-dimethylethyl[2-({[4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-3-fluorophenyl]carbonyl}amino)-3-phenylpropyl]carbamate

To a solution of 4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-3-fluorobenzoicacid (100 mg, 0.43 mmol), 1,1-dimethylethyl(2-amino-3-phenylpropyl)carbamate (108 mg, 0.43 mmol), diisopropylethylamine (0.4 mL, 2.14 mmol) in DCM (5 mL) was added PyBrop (330 mg, 0.71mmol) in one portion. After 1 h, the reaction contents were partitionedbetween H₂O/DCM. The aqueous phase was washed several times with DCM.The combined organic fractions were dried (Na₂SO₄), concentrated andpurified via column chromatography (silica) affording the title compound(79 mg, 38%) as a white solid: LCMS (ES) m/z=487 (M+H)+.

d)N-[2-amino-1-(phenylmethyl)ethyl]-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-3-fluorobenzamide

A solution of1,1-dimethylethyl[2-({[4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-3-fluorophenyl]carbonyl}amino)-3-phenylpropyl]carbamatein TFA-DCM (3 mL, 1:2) was stirred at 25° C. After 30 min, the solutionwas concentrated and the residue neutralized through a silica plug (4%MeOH in DCM (1% NH₄OH)) affording the free base of the title compound.

The free base, as a solution in MeOH, was then treated with excess 4MHCl in dioxane affording a white solid as the HCl salt of the titlecompound (17 mg, 17%): LC-MS (ES) m/z 387 (M+H)⁺, ¹H NMR (400 MHz, MeOD)δ ppm 3.01-3.07 (m, 2H) 3.18-3.29 (m, 2H) 3.73-3.79 (m, 3H) 4.61 (d,J=3.54 Hz, 1H) 7.26 (d, J=4.80 Hz, 1H) 7.34 (d, J=3.03 Hz, 4H) 7.57 (t,J=7.45 Hz, 1H) 7.60-7.66 (m, 1H) 7.71-7.80 (m, 2H).

Example 28

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-3-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to Example27, except substituting 4-bromo-3-chlorobenzoic acid (282 g, 1.2 mmol)for 4-bromo-3-fluorobenzoic acid: LC-MS (ES) m/z 403 (M+H)⁺, ¹H NMR (400MHz, MeOD) δ ppm 2.99-3.10 (m, 2H) 3.19-3.29 (m, 2H) 3.69 (s, 3H)4.59-4.67 (m, 1H) 7.22-7.28 (m, 1H) 7.30-7.36 (m, 4H) 7.52 (d, J=8.08Hz, 1H) 7.59 (s, 1H) 7.88 (ddd, J=7.96, 3.92, 1.77 Hz, 1H) 8.02 (dd,J=4.17, 1.64 Hz, 1H).

Example 29

Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluoro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)benzamidea). 3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzoic acid

To a solution of 4-bromo-3-fluorobenzoic acid (220 mg, 1.0 mmol) indioxane/H₂O (5:1, 12 mL) was added K₂CO₃ (414 mg, 3.0 mmol),tetrakistriphenylphosphine Pd(0) (58 mg, 0.05 mmol) and5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole (214 mg,1.1 mmol). The reaction mixture was heated to 80° C. in a sealed tubefor 12 h and was then partitioned between 6N NaOH and DCM. The pH of theaqueous phase was adjusted to ˜3 with 3M HCl and washed several timeswith DCM. The combined organic fractions were dried (Na₂SO₄),concentrated under vacuum and used directly without further purification(196 mg, 89%): LC-MS (ES) m/z=221 (M+H)⁺.

b). 4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-3-fluorobenzoic acid

A solution of 3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzoic acid (196 mg,0.89 mmol) and N-chlorosuccinimide (119 mg, 0.89 mmol) in THF (5 mL) wasstirred in a sealed tube at 70° C. After 1 h, the solution waspartitioned between H₂O-DCM, the aqueous phase was adjusted to pH 3 andthe aqueous phase was washed several times with DCM. The combinedorganic fractions were dried (Na₂SO₄), concentrated under vacuum andused directly without further purification (192 mg, 85%): LC-MS (ES)m/z=255 (M+H)⁺.

c).4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-N-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluorobenzamide

To a solution of 4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-3-fluorobenzoicacid (100 mg, 0.43 mmol),2-{(2S)-2-amino-3-[2-(trifluoromethyl)phenyl]propyl}-1H-isoindole-1,3(2H)-dione-HCl(151 mg, 0.43 mmol) [prepared according to Preparation 3],diisopropylethyl amine (0.4 mL, 2.14 mmol) in DCM (5 mL) was addedPyBrop (330 mg, 0.71 mmol) in one portion. After 1 h, the reactioncontents were partitioned between H₂O/DCM. The aqueous phase was washedseveral times with DCM. The combined organic fractions were dried(Na₂SO₄), concentrated and purified via column chromatography (silica)affording the title compound (90 mg, 36%) as a white solid: LCMS (ES)m/z=585 (M+H)+.

d)N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluoro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)benzamide

To a solution of4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-N-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluorobenzamide(90 mg, 0.15 mmol) in MeOH/THF (2 mL, 1:1) at RT was added hydrazine(0.12 mL, 3.93 mmol). After stirring for 18 h at RT, the reactionsolution was concentrated under vacuum and purified via columnchromatography (silica, 3% MeOH in DCM (1% NH4OH)) yielding the titlecompound (14 mg, 21%).

The neutral compound from above was dissolved in MeOH (2 mL), treatedwith excess 4M HCl in dioxane (0.5 mL) and concentrated affording theHCl salt of the title compound as a light yellow solid: LC-MS (ES) m/z455 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 3.17 (d, J=9.09 Hz, 1H) 3.26(d, J=6.32 Hz, 3H) 3.72-3.80 (m, 3H) 4.74 (s, 1H) 7.41-7.49 (m, 1H)7.54-7.64 (m, 4H) 7.72-7.84 (m, 3H).

Example 30

Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)benzamide

The title compound was prepared as a white solid according to Example29, except substituting 4-bromo-3-chlorobenzoic acid (282 mg, 1.2 mmol)for 4-bromo-3-fluorobenzoic acid: LC-MS (ES) m/z 471 (M+H)⁺, ¹H NMR (400MHz, MeOD) δ ppm 3.20 (d, J=9.35 Hz, 1H) 3.28 (d, J=3.54 Hz, 2H) 3.37(s, 1H) 3.70 (s, 3H) 4.75 (d, J=4.55 Hz, 1H) 7.41-7.47 (m, 1H) 7.55 (d,J=7.83 Hz, 2H) 7.59-7.62 (m, 2H) 7.72 (d, J=7.58 Hz, 1H) 7.96 (d, J=8.08Hz, 1H) 8.09 (s, 1H).

Example 31

Preparation ofN-[2-amino-1-(phenylmethyl)ethyl]-3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzamidea) methyl 3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzoate

To a solution of methyl 3-bromo-4-iodobenzoate (409 mg, 1.2 mmol) indioxane/H₂O (4:1, 10 mL) was added K₂CO₃ (497 mg, 3.6 mmol),tetrakistriphenylphosphine Pd(0) (69 mg, 0.06 mmol) and5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole (349 mg,3.6 mmol). The reaction mixture was heated to 80° C. in a sealed tubefor 12 h and was then partitioned between 6N NaOH and DCM. The pH of theaqueous phase was adjusted to ˜3 with 3M HCl and washed several timeswith DCM. The combined organic fractions were dried (Na₂SO₄),concentrated under vacuum and used directly without further purification(307 mg, 87%): LC-MS (ES) m/z=296 (M+H)+.

b) 3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzoic acid

To a solution of methyl 3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzoate(275 mg, 093 mmol) in MeOH (10 mL) was added NaOH solution (2.5 M, 3.72mL, 9.3 mmol). After 12 h at 25° C., the solution was partitionedbetween H₂O-DCM and the pH of the aqueous phase was adjusted to ˜4 with6N HCl solution. The aqueous phase was washed several times with DCM andthe combined organic fractions were dried over Na₂SO₄ and concentratedto afford the title compound as a white solid (230.1 mg, 88%): LC-MS(ES) m/z=282 (M+H)+.

c)1,1-dimethylethyl[2-({[3-bromo-4-(1-methyl-1H-pyrazol-5-yl)phenyl]carbonyl}amino)-3-phenylpropyl]carbamate

To a solution of 3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzoic acid (100mg, 0.36 mmol), PyBrOP (277 mg, 0.59 mmol) and diisopropylethyl amine(0.11 mL, 0.59 mmol) in DCM (6 mL) at 25° C. was added 1,1-dimethylethyl(2-amino-3-phenylpropyl)carbamate (90 mg, 0.36 mmol)[prepared inPreparation 2]. After 16 h, the solution was concentrated and purifiedvia column chromatography (silica, hexanes/EtOAc, 2:1) affording thetitle compound (87 mg, 47%) as a white solid: LC-MS (ES) m/z=514 (M+H)⁺.

d)N-[2-amino-1-(phenylmethyl)ethyl]-3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzamide

1,1-dimethylethyl[2-({[3-bromo-4-(1-methyl-1H-pyrazol-5-yl)phenyl]carbonyl}amino)-3-phenylpropyl]carbamate(87 g, 0.17 mmol) was dissolved in MeOH (1 mL) and CHCl₃ (10 mL) andtreated with 4M HCl in dioxane (5 mL). After 3 h, the solution wasconcentrated and purified via column chromatography (silica, CHCl₃/MeOH,90:10 containing 1% NH₄OH affording the free base of the title compound(30.7 mg, 44%) as a white powder.

The free base, as a solution in MeOH (2 mL) was treated with an excessof 1M HCl in Et₂O affording the final compound as the HCl salt: LC-MS(ES) m/z 413(M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 2.98-3.08 (m, 2H)3.14-3.29 (s, 2H) 3.70 (s, 3H) 4.63 (s, 1H) 6.38 (s, 1H) 7.25 (s, 1H)7.33 (s, 4H) 7.50 (s, 1H) 7.60 (s, 1H) 7.84 (d, J=7.83 Hz, 1H) 8.12 (s,1H).

Example 32

Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzamidea) methyl 3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzoate

To a solution of methyl 3-bromo-4-iodobenzoate (409 mg, 1.2 mmol) indioxane/H₂O (4:1, 10 mL) was added K₂CO₃ (497 mg, 3.6 mmol),tetrakistriphenylphosphine Pd(0) (69 mg, 0.06 mmol) and5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole (349 mg,3.6 mmol). The reaction mixture was heated to 80° C. in a sealed tubefor 12 h and was then partitioned between 6N NaOH and DCM. The pH of theaqueous phase was adjusted to ˜3 with 3M HCl and washed several timeswith DCM. The combined organic fractions were dried (Na₂SO₄),concentrated under vacuum and used directly without further purification(307 mg, 87%): LC-MS (ES) m/z=296 (M+H)⁺.

b) 3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzoic acid

To a solution of methyl 3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzoate(275 mg, 0.93 mmol) in MeOH (10 mL) was added NaOH solution (2.5 M, 3.72mL, 9.3 mmol). After 12 h at 25° C., the solution was partitionedbetween H₂O-DCM and the pH of the aqueous phase was adjusted to ˜4 with6N HCl solution. The aqueous phase was washed several times with DCM andthe combined organic fractions were dried over Na₂SO₄ and concentratedto afford the title compound as a white solid (230.1 mg, 88%): LC-MS(ES) m/z=282 (M+H)⁺.

c)3-bromo-N-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-pyrazol-5-yl)benzamide

To a solution of 3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzoic acid (100mg, 0.36 mmol), PyBrOP (277 mg, 0.59 mmol) and diisopropylethyl amine(0.11 mL, 0.59 mmol) in DCM (6 mL) at 25° C. was added2-{(2S)-2-amino-3-[2-(trifluoromethyl)phenyl]propyl}-1H-isoindole-1,3(2H)-dione(161 mg, 0.36 mmol) [prepared according to Preparation 3]. After 16 h,the solution was concentrated and purified via column chromatography(silica, hexanes/EtOAc, 2:1) affording the title compound (172 mg, 78%)as a white solid: LC-MS (ES) m/z=612 (M+H)⁺.

d)N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzamide

To a solution of3-bromo-N-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-pyrazol-5-yl)benzamide(172 mg, 0.28 mmol) in MeOH/THF (4 mL, 1:1) at RT was added hydrazine(0.17 mL, 5.5 mmol). After stirring for 18 h at RT, the reactionsolution was concentrated under vacuum and purified via columnchromatography (silica, 3% MeOH in DCM (1% NH₄OH)) yielding the titlecompound (60.4 mg, 45%).

The neutral compound from above was dissolved in MeOH (2 mL), treatedwith excess 4M HCl in dioxane (500 μL) and concentrated affording theHCl salt of the title compound as a white solid: LC-MS (ES) m/z 482(M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 3.09-3.19 (m, 1H) 3.26 (d, J=6.32Hz, 3H) 3.74 (s, 3H) 4.73 (s, 1H) 6.45 (s, 1H) 7.46 (s, 1H) 7.55 (t,J=3.79 Hz, 3H) 7.73 (d, J=7.83 Hz, 2H) 7.92 (s, 1H) 8.22 (s, 1H).

Example 33

Preparation of3-amino-N-[3-chloro-4-(1-methyl-1H-pyrazol-5-yl)phenyl]-2-phenylpropanamidea)1,1-dimethylethyl{3-[(4-bromo-3-chlorophenyl)amino]-3-oxo-2-phenylpropyl}carbamate

To a solution of3-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-phenylpropanoic acid (159mg, 0.6 mmol), PyBrOP (462 mg, 0.99 mmol) and diisopropylethyl amine(0.35 mL, 2.0 mmol) in DCM (6 mL) at 25° C. was added4-bromo-3-chloroaniline (124 mg, 0.6 mmol). After 16 h, the solution wasconcentrated and purified via column chromatography (silica,hexanes/EtOAc, 2:1) affording the title compound (248 mg, 91%) as awhite solid: LC-MS (ES) m/z=454 (M+H)⁺.

b)1,1-dimethylethyl(3-{[3-chloro-4-(1,3-dimethyl-1H-pyrrol-2-yl)phenyl]amino}-3-oxo-2-phenylpropyl)carbamate

To a solution of1,1-dimethylethyl(3-{[3-chloro-4-(1-methyl-1H-pyrrol-2-yl)phenyl]amino}-3-oxo-2-phenylpropyl)carbamate(247 mg, 0.54 mmol) in dioxane/H₂O (4:1, 10 mL) was added K₂CO₃ (224 mg,1.62 mmol), tetrakistriphenylphosphine Pd(0) (31 mg, 0.03 mmol) and5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methyl-1H-pyrazole (157 mg,1.62 mmol). The reaction mixture was heated to 80° C. in a sealed tubefor 12 h and was then partitioned between H—₂O-DCM. The aqueous phasewas washed several times with DCM and the combined organic fractionswere dried (Na₂SO₄), concentrated under vacuum and used directly withoutfurther purification (228 mg, 93%): LC-MS (ES) m/z=455 (M+H)⁺.

c)3-amino-N-[3-chloro-4-(1-methyl-1H-pyrazol-5-yl)phenyl]-2-phenylpropanamide

1,1-dimethylethyl(3-{[3-chloro-4-(1,3-dimethyl-1H-pyrrol-2-yl)phenyl]amino}-3-oxo-2-phenylpropyl)carbamate(228 mg, 0.5 mmol) was dissolved in MeOH (1 mL) and CHCl₃ (10 mL) andtreated with 4M HCl in dioxane (5 mL). After 3 h, the solution wasconcentrated and purified via column chromatography (silica, CHCl₃/MeOH,90:10 containing 1% NH₄OH) affording the free base of the title compound(146.4 mg, 83%) as a white powder.

The free base, as a solution in MeOH was treated with an excess of 1MHCl in Et₂O to yield the final compound as the HCl salt: LC-MS (ES) m/z355 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 3.28 (dd, J=12.88, 5.31 Hz, 1H)3.59-3.70 (m, 1 H) 3.87 (d, J=11.37 Hz, 3H) 4.22 (d, J=2.27 Hz, 1H) 6.67(s, 1H) 7.39-7.50 (m, 6 H) 7.67 (s, 1H) 8.14 (d, J=5.05 Hz, 2H).

Example 34

Preparation of3-amino-N-[3-chloro-4-(1-methyl-1H-pyrazol-5-yl)phenyl]-2-(phenylmethyl)propanamide

The title compound was prepared as a white solid according to Example33, except substituting3-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-(phenylmethyl)propanoicacid (168 mg, 0.6 mmol)[prepared according to Preparation 5] for3-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-phenylpropanoic acid:LC-MS (ES) m/z 355 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 2.97 (dd,J=13.52, 7.20 Hz, 1H) 3.07 (dd, J=12.51, 3.41 Hz, 1H) 3.11-3.18 (m, 1H)3.27 (s, 1H) 3.34-3.39 (m, 1H) 3.88 (s, 3H) 6.71 (s, 1H) 7.25 (d, J=4.29Hz, 1H) 7.32 (d, J=4.04 Hz, 4H) 7.46 (d, J=8.59 Hz, 1H) 7.62-7.66 (m,1H) 8.08 (d, J=1.77 Hz, 1H) 8.11 (s, 1H).

Example 35 Preparation of3-amino-N-[3-(1-methyl-1H-pyrazol-5-yl)phenyl]-2-(phenylmethyl)propanamide

a)1,1-dimethylethyl[3-[(3-bromophenyl)amino]-3-oxo-2-(phenylmethyl)propyl]carbamate

To a solution of3-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2-(phenylmethyl)propanoicacid (360 mg, 1.29 mmol), PyBrOP (900 mg, 1.93 mmol) anddiisopropylethyl amine (0.9 mL, 5 mmol) in DCM (10 mL) at 25° C. wasadded 3-bromoaniline (0.29 mg, 1.66 mmol). After 2 h, the solution wasconcentrated and purified via column chromatography (silica,hexanes/EtOAc, 10:1 to 3:1) affording the title compound (400 mg, 72%)as a white solid: LC-MS (ES) m/z=434 (M+H)⁺.

b)1,1-dimethylethyl[3-{[3-(1-methyl-1H-pyrazol-5-yl)phenyl]amino}-3-oxo-2-(phenylmethyl)propyl]carbamate

To a solution of1,1-dimethylethyl[3-[(3-bromophenyl)amino]-3-oxo-2-(phenylmethyl)propyl]carbamate(230 mg, 0.53 mmol) in dioxane/H₂O (5:1, 6 mL) was added K₂CO₃ (220 mg,1.59 mmol), tetrakistriphenylphosphine Pd(0) (61 mg, 52 μmol) and1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(164 mg, 0.79 mmol). The reaction mixture was heated to 70° C. in asealed tube. After 10 h, the reaction mixture was concentrated undervacuum and purified on silica (hexanes/EtOAc, 30-60%) to afford thetitle compound (0.17 g, 74%) as a light yellow solid: LC-MS (ES) m/z=435(M+H)⁺.

c)3-amino-N-[3-(1-methyl-1H-pyrazol-5-yl)phenyl]-2-(phenylmethyl)propanamide

1,1-dimethylethyl[3-{[3-(1-methyl-1H-pyrazol-5-yl)phenyl]amino}-3-oxo-2-(phenylmethyl)propyl]carbamate(170 mg, 0.39 mmol) was dissolved in DCM (10 mL) and treated with TFA (1mL). After 1 h, the solution was concentrated and purified usingreverse-phase HPLC (C18 column: H₂O/CH₃CN, 40-10%) affording the bis-TFAsalt of the title compound (115 mg, 52%) as a white solid. LC-MS (ES)m/z=335 (M+H)⁺, ¹H NMR (d4-MeOD, 400 MHz) δ 7.76 (br s, 1H), 7.43 (br s,1 h), 7.35-7.23 (m, 9H), 4.55 (m, 1H), 3.72 (s, 3H), 3.23 (dd, J=3.5,13.1 Hz, 1H), 3.13 (dd, J=10.4, 13.1 Hz, 1H), and 3.03-2.95 (m, 2H).

Example 36 Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide

a) 4-(1-methyl-1H-1,2,4-triazol-5-yl)benzoic acid

A mixture of 5-iodo-1-methyl-1H-1,2,4-triazole (418 mg, 2.00 mmol),{4-[(ethyloxy)carbonyl]phenyl}boronic acid (466 mg, 2.40 mmol), andBIS(TRIPHENYLPHOSPHINE)PALLADIUM(II) CHLORIDE (70.2 mg, 0.10 mmol) intetrahydrofuran (THF) (10 mL) was stirred under N₂ in a 25 mL microwavereaction vessel. Sodium carbonate (1000 μl, 2.00 mmol) was added. Thereaction mixture was purged with N₂, sealed and irradiated with MW at138° C. for 28 min. The crude reaction mixture was loaded directly on a25M biotage column, which was eluted with 50-100% of EtOAc/hexane togive the ester intermediate. This ester was hydrolyzed with 2N NaOH (4mL) in THF (8 mL) at 70° C. The reaction was completed in one hour.

The organic layer was washed with H₂O 3× and discarded. The combinedaqueous layers were acidified with 2N HCl. The product had poorsolubility, and it was extracted with MeOH/CHCl₃ 5×. The combinedextracts were dried, and concentrated to give 0.36 g of light yellowsolid (89%). LC-MS (ES) m/z=204 (M+H)⁺.

b)N-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide

Hunig's base (0.084 ml, 0.48 mmol) was added to a suspension of4-(1-methyl-1H-1,2,4-triazol-5-yl)benzoic acid (39.3 mg, 0.19 mmol),2-{(2S)-2-amino-3-[2-(trifluoromethyl)phenyl]propyl}-1H-isoindole-1,3(2H)-dione(62 mg, 0.16 mmol), and PyBroP (75 mg, 0.16 mmol) in DCM (1 ml). Thereaction mixture became clear and was stirred at RT overnight. Thereaction mixture was loaded directly on a 25S Biotage column, which waseluted with 50-100% EA/DCM to give 67.3 mg of product. LC-MS (ES)m/z=534 (M+H)⁺.

c)N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide

Hydrazine hydrate (0.028 ml, 0.58 mmol) was added to a solution ofN-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide(67.3 mg, 0.116 mmol) in DCM/MeOH (1/0.1 ml).

The reaction mixture was stirred at RT overnight. A white precipitateformed. The reaction mixture was filtered through a syringe filter,concentrated and purified by RP-HPLC. The fractions containing productwere concentrated and treated with 2N HCl aq. solution 3× to giveproduct (19.3 mg, 36%) as a white solid. LC-MS (ES) m/z=404 (M+H)⁺, ¹HNMR (d4-MeOD, 400 MHz) δ 8.74 (s, 1H), 8.13 (m, 2H), 7.95 (m, 2H), 7.72(m, 1H), 7.60-7.51 (m, 2H), 7.44 (m, 1H), 4.71 (m, 1H), 4.15 (s, 3H),3.35-3.15 (m, 4H).

Example 37

Preparation ofN-[(1S)-2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide

The title compound was prepared as a white solid according to Example36, except substituting2-[(2S)-2-amino-3-phenylpropyl]-1H-isoindole-1,3(2H)-dione [preparedaccording to Preparation 3] for2-{(2S)-2-amino-3-[2-(trifluoromethyl)phenyl]propyl}-1H-isoindole-1,3(2H)-dione:LC-MS (ES) m/z 336 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 8.82 (s, 1H),8.09-8.07 (m, 2H), 7.95-7.92 (m, 2H), 7.35-7.23 (m, 5H), 4.65 (m, 1H),4.15 (s, 3H), 3.3-3.0 (m, 4H).

Example 38

Preparation ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide

The title compound was prepared as a white solid according to Example36, except substituting2-[(2S)-2-amino-3-(3-fluorophenyl)propyl]-1H-isoindole-1,3(2H)-dione[prepared according to Preparation 3] for2-{(2S)-2-amino-3-[2-(trifluoromethyl)phenyl]propyl}-1H-isoindole-1,3(2H)-dione:LC-MS (ES) m/z 354 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 8.91 (s, 1H),8.12-8.10 (m, 2H), 7.31 (m, 1H), 7.18-7.10 (m, 2H), 6.96 (m, 1H), 4.65(m, 1H), 4.16 (s, 3H), 3.3-3.0 (m, 4H).

Example 39

Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide

The title compound was prepared as a white solid according to Example36, except substituting {2-chloro-4-[(methyloxy)carbonyl]phenyl}boronicacid for {4-[(ethyloxy)carbonyl]phenyl}boronic acid: LC-MS (ES) m/z404.2 (M+H)⁺, ¹H NMR (d₄-MeOH, 400 MHz) δ 8.38 (m, 1H), 8.11 (m, 1H),7.96 (m, 1H), 7.73 (m, 2H), 7.56 (m, 2H), 7.45 (m, 1H), 4.76 (m, 1H),3.88 (s, 3H), 3.26 (m, 2H), 3.16 (m, 2H).

Example 40

Preparation ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide

The title compound was prepared as a white solid according to Example38, except substituting {2-chloro-4-[(methyloxy)carbonyl]phenyl}boronicacid for {4-[(ethyloxy)carbonyl]phenyl}boronic acid: LC-MS (ES) m/z388.2 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 8.13 (m, 1H) 8.01 (s, 1H)7.87 (d, 1H), 7.66 (d, 1H) 7.33 (m, 1H) 7.15-6.98 (m, 3H) 4.65 (m, 1H)3.80 (s, 3H) 3.23 (m, 2H) 3.05 (m, 2H).

Example 41

Preparation ofN-[(1S)-2-amino-1-(phenylmethyl)ethyl]-3-chloro-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide

The title compound was prepared as a white solid according to Example37, except substituting {2-chloro-4-[(methyloxy)carbonyl]phenyl}boronicacid for {4-[(ethyloxy)carbonyl]phenyl}boronic acid: LC-MS (ES) m/z 370(M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 8.65 (s, 1H), 8.09 (d, J=1 Hz, 1H),7.96 (dd, J=8, 1 Hz, 1H), 7.79 (d, J=8 Hz, 1H), 7.34-7.24 (m, 5H), 4.62(m, 1H), 4.16 (s, 3H), 3.3-3.0 (m, 4H).

Example 42 Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide

a) 4-(1-methyl-1H-1,2,3-triazol-5-yl)benzoic acid

Triethylamine (0.28 ml, 2.00 mmol) was added to a mixture of1-methyl-5-(tributylstannanyl)-1H-1,2,3-triazole (781 mg, 2.1 mmol),methyl 4-bromobenzoate (430 mg, 2.0 mmol), andBIS(TRIPHENYLPHOSPHINE)PALLADIUM(II) CHLORIDE (70.2 mg, 0.10 mmol) intoluene (10 ml) to give a yellow suspension. The reaction mixture waspurged with N₂, sealed and stirred at 100° C. for 3 hr. Potassiumfluoride (465 mg, 8.00 mmol) and MeOH (2 mL) were added. The mixture wasstirred at RT for 2 h, filtered on celite, and rinsed with EtOAc. Thecombined filtrates were concentrated and the residue was purified on a25M biotage column to give 0.5 g of methyl4-(1-methyl-1H-1,2,3-triazol-5-yl)benzoate as a tan solid.

A mixture of methyl 4-(1-methyl-1H-1,2,3-triazol-5-yl)benzoate (0.5 g,2.30 mmol) and Sodium hydroxide (4.60 ml, 9.21 mmol, 2N aqueoussolution) in tetrahydrofuran (THF) was heated at 70° C. for 1 hr. Theorganic layer was washed with H₂O 3× and discarded. The combined aqueouslayers were acidified with 2N HCl aqueous solution. The productprecipitated and was filtered, rinsed with H₂O and dried under vacuum togive 0.3 g (74%) as a tan solid. LC-MS (ES) m/z=204 (M+H)⁺.

b)N-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide

The title compound was prepared according to procedure 36(b) exceptsubstituting 4-(1-methyl-1H-1,2,3-triazol-5-yl)benzoic acid for4-(1-methyl-1H-1,2,4-triazol-5-yl)benzoic acid: LC-MS (ES) m/z=534(M+H)⁺.

c)N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide

The title compound was prepared according to procedure 36(c) exceptsubstitutingN-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamideforN-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,4-triazol-5-yl)benzamide:LC-MS (ES) m/z=404 (M+H)⁺, ¹H NMR (d4-MeOD, 400 MHz) δ 8.34 (s, 1H),8.05-8.02 (m, 2H), 7.78-7.71 (m, 3H), 7.58-7.52 (m, 2H), 7.43 (m, 1H),4.73 (m, 1H), 4.26 (s, 3H), 3.3-3.1 (m, 4H).

Example 43 Preparation ofN-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(4-chloro-1-methyl-1H-1,2,3-triazol-5-yl)benzamide

A mixture ofN-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide(0.1 g, 0.16 mmol) and NCS (0.043 g, 0.32 mmol) in DMF (1 mL) was heatedat 50° C. for 2 hr. The reaction mixture was diluted with EtOAc, washedwith H₂O 3×, dried and concentrated to give the crude product.

The crude chlorotriazole was dissolved in MeOH (0.1 mL) and DCM (1 mL).Hydrazine hydrate (0.04 ml, 0.80 mmol) was added. The reaction mixturewas stirred at RT overnight. A white precipitate formed and was filteredthrough a syringe filter. The filtrates were concentrated and theresidue was purified by RP-HPLC. The product fractions were concentratedand treated with 2N HCl aq., which was concentrated to give 65.8 mg ofproduct as a light yellow solid. LC-MS (ES) m/z=438.2 (M+H)⁺, ¹H NMR(d4-MeOD, 400 MHz) δ ppm 8.34 (s, 1H), 8.02 (m, 2H), 7.74-7.68 (m, 3H),7.56 (m, 2H), 7.43 (m, 1H), 4.73 (m, 1H), 4.08 (s, 3H), 3.36-3.12 (m,4H).

Example 44 Preparation ofN-[(1S)-2-amino-1-(phenylmethyl)ethyl]-4-(4-chloro-1-methyl-1H-1,2,3-triazol-5-yl)benzamide

The title compound was prepared as a white solid according to theprocedure of Example 43, except substitutingN-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-[(2-phenyl)methyl]ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamideforN-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide:LC-MS (ES) m/z 370.2 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 7.98 (m, 2),7.66 (m, 2H), 7.34-7.23 (m, 5H), 7.33 (m, 1H), 4.63 (m, 1H), 4.07 (s,3H) 3.3-3.0 (m, 4H).

Example 45 Preparation ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-4-(4-chloro-1-methyl-1H-1,2,3-triazol-5-yl)benzamide

The title compound was prepared as a white solid according to theprocedure of Example 43, except substitutingN-{(1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-[(3-fluorophenyl)methyl]ethyl}-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamideforN-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide.The title compound was a TFA salt: LC-MS (ES) m/z 388.2 (M+H)⁺, ¹H NMR(400 MHz, MeOD) δ ppm 7.95 (m, 2H), 7.68 (m, 2H), 7.34 (m, 1H),7.15-7.07 (m, 2H), 6.98 (m, 1H), 4.63 (m, 1H), 4.07 (s, 3H), 3.29-2.95(m, 4H).

Example 46 Preparation ofN-[(1S)-2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide

The title compound was prepared as a white solid according to theprocedure of Example 42, except substitutingN-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-[(2-phenyl)methyl]ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamideforN-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide:LC-MS (ES) m/z 336.2 (M+H)⁺, ¹H NMR (400 MHz, MeOD) δ ppm 8.65 (s, 1H),8.02 (m, 2H), 7.80 (m, 2H), 7.36-7.22 (m, 5H), 4.64 (m, 1H), 4.32 (s,3H), 3.3-3.0 (m, 4H).

Example 47 Preparation ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide

The title compound was prepared as a white solid according to theprocedure of Example 43, except substitutingN-{(1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-[(3-fluorophenyl)methyl]ethyl}-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamideforN-((1S)-2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-(1-methyl-1H-1,2,3-triazol-5-yl)benzamide.The title compound was a TFA salt: LC-MS (ES) m/z 354.2 (M+H)⁺, ¹H NMR(400 MHz, MeOD) δ ppm 7.94-7.90 (m, 3H), 7.69 (m, 2H), 7.33 (m, 1H)7.15-7.07 (m, 2H), 6.98 (m, 1H), 4.62 (m, 1H), 4.15 (s, 3H), 3.30-2.96(m, 4H).

Example 48 Capsule Composition

An oral dosage form for administering the present invention is producedby filing a standard two piece hard gelatin capsule with the ingredientsin the proportions shown in Table I, below.

TABLE I INGREDIENTS AMOUNTSN-[2-amino-1-(phenylmethyl)ethyl]-5-(1-methyl- 25 mg1H-pyrazol-5-yl)-2-pyridinecarboxamide (Compound of Example 1) Lactose55 mg Talc 16 mg Magnesium Stearate  4 mg

Example 49 Injectable Parenteral Composition

An injectable form for administering the present invention is producedby stirring 1.5% by weight ofN-[2-amino-1-(phenylmethyl)ethyl]-6-(1-methyl-1H-pyrazol-5-yl)-3-pyridinecarboxamide(compound of Example 2), in 10% by volume propylene glycol in water.

Example 50 Tablet Composition

The sucrose, calcium sulfate dihydrate and an Akt inhibitor as shown inTable II below, are mixed and granulated in the proportions shown with a10% gelatin solution. The wet granules are screened, dried, mixed withthe starch, talc and stearic acid;, screened and compressed into atablet.

TABLE II INGREDIENTS AMOUNTSN-(2-amino-1-benzylethyl)-5-(1-methyl-1H-pyrazol-5- 20 mg yl)pyrimidine-2-carboxamide (Compound of Example 3) calcium sulfatedehydrate 30 mg  Sucrose 4 mg Starch 2 mg Talc 1 mg stearic acid 0.5mg  

While the preferred embodiments of the invention are illustrated by theabove, it is to be understood that the invention is not limited to theprecise instructions herein disclosed and that the right to allmodifications coming within the scope of the following claims isreserved.

1. A compound of Formula (I):

wherein: Q is selected from: phenyl, substituted phenyl, benzyl, andbenzyl wherein the aromatic ring is substituted; R¹ is selected from:hydrogen, trifluoromethyl, hydroxy, —C₁-C₂alkyl, and halogen; L is—C(H)—; P is C(R⁴⁰)—, where R⁴⁰ is selected from: hydrogen, —C₁-C₄alkyl,and halogen; A is selected from: —C(O)— and —N(H)—; B is selected from:—C(O)— and —N(H)—; and X, Y and Z are independently selected from:—C(H)—, and —C(R²)—, wherein R² is selected from halogen,trifluoromethyl, hydroxy, and —C₁-C₄alkyl; or a pharmaceuticallyacceptable salt thereof; provided: A and B are not the same.
 2. Acompound of claim 1 selected from:N-[2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-pyrazol-5-yl)benzamide;N-[2-amino-1-(phenylmethyl)ethyl]-2-chloro-4-(1-methyl-1H-pyrazol-5-yl)benzamide;N-[2-amino-1-(phenylmethyl)ethyl]-3-chloro-4-(1-methyl-1H-pyrazol-5-yl)benzamide;N-[2-amino-1-(phenylmethyl)ethyl]-4-(1-methyl-1H-pyrazol-5-yl)-3-(trifluoromethyl)benzamide;N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-chloro-4-(1-methyl-1H-pyrazol-5-yl)benzamide;N-[2-amino-1-(phenylmethyl)ethyl]-3-methyl-4-(1-methyl-1H-pyrazol-5-yl)benzamide;N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzamide;N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-methyl-4-(1-methyl-1H-pyrazol-5-yl)benzamide;N-[2-amino-1-(phenylmethyl)ethyl]-3-hydroxy-4-(1-methyl-1H-pyrazol-5-yl)benzamide;N-[2-amino-1-(phenylmethyl)ethyl]-3-fluoro-4-(1-methyl-1H-pyrazol-5-yl)benzamide;N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-4-chloro-5-(1-methyl-1H-pyrazol-5-yl)-2-pyridinecarboxamide;N-[2-amino-1-(phenylmethyl)ethyl]-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-3-fluorobenzamide;N-[2-amino-1-(phenylmethyl)ethyl]-3-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)benzamide;N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-fluoro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)benzamide;N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)benzamide;N-[2-amino-1-(phenylmethyl)ethyl]-3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzamide;N-((1S)-2-amino-1-{[2-(trifluoromethyl)phenyl]methyl}ethyl)-3-bromo-4-(1-methyl-1H-pyrazol-5-yl)benzamide;3-amino-N-[3-chloro-4-(1-methyl-1H-pyrazol-5-yl)phenyl]-2-phenylpropanamide;and3-amino-N-[3-chloro-4-(1-methyl-1H-pyrazol-5-yl)phenyl]-2-(phenylmethyl)propanamide;or pharmaceutically acceptable salt thereof.
 3. A pharmaceuticalcomposition comprising a compound according to claim 1 and apharmaceutically acceptable carrier.